LANCE Ultra Substrate Selection Guide

FINDING THE
PATHWAY TO
ASSAY OPTIMIZATION
IS EASY
LANCE® Ultra Kinase Assay Selection Guide
LANCE Ultra Serine/Threonine Kinase Selection Guide
LANCE® Ultra TR-FRET reagents comprise the widest portfolio of validated kinase assay offerings available for rapid, sensitive
and robust screening of purified kinase targets in a biochemical format.
• We provide S/B ratiometric data for each LANCE Ultra assay to guide you to
the best performing solution for your assay.
• Rapid assay optimization every time.
• Our selection guides contain over 300 kinases from a variety of suppliers:
– 225 Serine/Threonine kinases validated on LANCE Ultra reagents
– 85 Tyrosine kinases validated on LANCE Ultra reagents
How to use this guide:
1.Locate your kinase
• In many cases, up to three commercial kinase vendors have been tested.
• Many common aliases are shown in parenthesis.
2.Best performing ULight ™ substrates are listed for each enzyme according
to performance
• Signal to background (S/B) ratios (Signal at 665 nm / minus ATP control
at 665 nm) are indicated in parenthesis.
• All S/B ratios were obtained at fixed experimental conditions unless
otherwise noted (see page 10).
If you cannot find your kinase of interest, please ask your PerkinElmer sales
specialist, as our list continues to expand. Two kits are available for
testing purposes:
• KinaSelect Ser/Thr kit (5 x 250 data points, TRF0300-C)
– 5 ULight-labeled Ser/Thr kinase specific substrates + 5 matching
Europium-labeled anti-phospho antibodies
• KinaSelect TK kit (1,000 data points, TRF0301-D)
– 1 ULight-labeled kinase specific substrate + 1 matching
Europium-labeled anti-phospho antibody
3.Based on your substrate choice, find the corresponding Europium-labeled
anti-phospho antibody on page 11 (i.e. LANCE Ultra products used for
this study) to identify the TR-FRET donor anti-phospho antibody for your
LANCE Ultra assay.
2
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LANCE® Ultra Reagents for Ser/Thr Kinase Assays
Up to three kinase substrates are listed based on signal to background (S/B) ratios obtained with commercially available enzymes.
S/B ratios (Signal at 665 nm / minus ATP control at 665 nm) are indicated in parenthesis.
Kinase ULight Substrate 1
ULight Substrate 2 ULight Substrate 3 ACTR2B (ACVR2B)ULight-Histone H3 (Thr3; 10.4)
ULight-Topo IIa (7.2)
ACV-R1 (ALK2)ULight-Topo IIa (10.9)
ALK2 (ACV-R1)ULight-Topo IIa (Mn2+; 21.5)
ACVR2AULight-Histone H3 (Thr3; 9.4)
ULight-CREBtide (5.7)
ACVRL1 (ALK1)ULight-CREBtide (14.2)
ULight-Topo IIa (5.6)
ULight-PLK (5.4)
BARK1 (ADRBK1, GRK2)ULight-Topo IIa (22.9)
ADRBK2 (BARK2, GRK3)ULight-Topo IIa (27.9)
BARK2 (ADRBK2, GRK3)ULight-Topo IIa (22.9)
ULight-Histone H3 (Thr3; 10.1)
AKT1ULight-CREBtide (32.0)
ULight-rpS6 (20.5)
ULight-PLK (19.1)
AKT2ULight-CREBtide (32.4) ULight-rpS6 (21.7)
ULight-PLK (20.1)
AKT3ULight-CREBtide (32.1) ULight-rpS6 (21.6)
ULight-PLK (20.7)
ALK4 (ACVR1B)ULight-Topo IIa (20.6)
AMPK-alpha1 flULight-CREBtide (9.6)
ULight-Histone H3 (Thr3; 5.3)
AMPKa1/b1/g1 (PRKAA1/B1/G1)ULight-CREBtide (31.9)
ULight-Acetyl-CoA Carboxylase (30.5) ULight-PLK (15.1)
AMPKa2/b1/g1(PRKAA2/B1/G1) ULight-CREBtide (30.3)
ULight-Acetyl-CoA Carboxylase (29.9)
ULight-PLK (11.5)
ARK5 (NuaK1)ULight-CREBtide (18.8)
ULight-Topo IIa (7.7)
NuaK1 (ARK5)ULight-CREBtide (22.8)
ULight-Topo IIa (3.1)
AurA (Aurora-A)ULight-PLK (20.5)
ULight-Histone H3 (Ser10; 13.0) AurB (AURKB)/INCENPULight-CREBtide (29.8)
ULight-rpS6 (21.0)
ULight-PLK (17.6)
Aurora-BULight-CREBtide (13.4)
ULight-Histone H3 (Ser10; 12.7) ULight-PLK (7.6)
AurC (AURKC)ULight-PLK (16.2)
ULight-Histone H3 (Ser10; 12.1) ULight-CREBtide (11.7)
Aurora-CULight-CREBtide (8.9)
ULight-Histone H3 (Ser10; 8.4) ULight-PLK (3.8)
BMPR1BULight-Topo IIa (18.7)
BMPR2ULight-Histone H3 (Thr3; 11.0)
BRAF
GST-MAP2K1/ ULight anti-GST (6.1)
BRAF [V600E]
GST-MAP2K1/ ULight anti-GST (10.5)
BRSK1ULight-CDC25 (7.6)
ULight-Acetyl-CoA Carboxylase (5.2) BRSK2ULight-CDC25 (8.4)
ULight-Acetyl-CoA Carboxylase (7.9) CaMK1a (CAMK1)2ULight-CREBtide (28.4)
CaMK1b (PNCK)2ULight-CREBtide (29.6)
CaMK1d (CAMK1D)2ULight-CREBtide (30.5)
CaMK2a (CAMK2A)2ULight-CREBtide (26.1)
ULight-rpS6 (22.3)
ULight-PKC (16.1)
CaMK2b (CAMK2B)2ULight-CREBtide (30.7)
ULight-rpS6 (23.6)
ULight-mTOR (12.0)
CaMK2d (CAMK2D)2ULight-rpS6 (25.4)
ULight-CREBtide (23.8)
ULight-PLK (14.0)
CaMK2g (CAMK2G)2ULight-CREBtide (26.2)
ULight-rpS6 (18.2)
ULight-PKC (15.6)
Kinase Supplier
Supplier Cat. No.
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ProQinase
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Invitrogen
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Invitrogen
Carna Biosciences
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Carna Biosciences
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ProQinase
Carna Biosciences
Carna Biosciences
ProQinase
Carna Biosciences
Carna Biosciences
Carna Biosciences
ProQinase
Carna Biosciences
ProQinase
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
09-132
0372-0000-1
09-134
09-131
PV4883
01-111
PV3827
01-112
01-101
01-102
01-103
09-135
0289-0000-1
02-113
02-114
0300-0000-1
02-126
05-101
05-102
0190-0000-1
05-103
0315-0000-1
09-138
09-139
09-122
09-144
02-115
02-116
02-104
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
02-105
02-106
02-109
02-110
02-111
02-112
Newly added Kinases highlighted in Green
3
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Kinase ULight Substrate 1
ULight Substrate 2 ULight Substrate 3
CaMK4 ULight-CREBtide (33.5)
ULight-mTOR (8.6)
ULight-PLK (6.0)
CAMKK12ULight-p70S6K (25.1)
ULight-CREBtide (19.6)
CAMKK2ULight-Histone H3 (Thr3; 12.2)
CAMKK22ULight-Histone H3 (Thr3; 12.0)
ULight-p70S6K (5.5)
CDC2/CycB1ULight-MBP (43.2)
ULight-4E-BP1 (37.2)
CDC2L6/CycCULight-Histone H3 (Thr3; 10.0)
CDC42BPB (MRCKb)ULight-Histone H3 (Ser10; 6.7) MRCKb (CDC42BPB)ULight-Histone H3 (Ser10; 6.9)
ULight-CREBtide (6.3) ULight-PLK (3.3)
CDC7/ASKULight-Topo IIa (14.6)
CDK1/CycAULight-4E-BP1 (37.9)
ULight-MBP (21.9)
CDK1/CycB1ULight-4E-BP1 (39.6)
ULight-MBP (27.8)
CDK1/CycEULight-4E-BP1 (39.2)
ULight-MBP (21.5)
CDK2/CycAULight-MBP (39.9)
ULight-4E-BP1 (36.7)
CDK2/CycE1ULight-4E-BP1 (33.5)
ULight-MBP (28.5)
CDK3/CycE1ULight-MBP (38.9)
ULight-4E-BP1 (23.3)
CDK4/CycD1ULight-4E-BP1 (38.7)
ULight-MBP (21.6)
CDK4/CycD3ULight-MBP (37.2)
ULight-4E-BP1 (32.3)
CDK5/p25ULight-MBP (37.1)
ULight-4E-BP1 (36.5)
CDK5/p25NCKULight-4E-BP1 (34.2)
ULight-MBP (24.9)
CDK5/p35NCKULight-4E-BP1 (40.3)
ULight-MBP (22.9)
CDK6/CycD1ULight-4E-BP1 (39.6)
ULight-MBP (17.0)
CDK6/CycD3ULight-MBP (35.7)
ULight-4E-BP1 (35.4)
CDK7/CycH/MAT1ULight-MBP (19.0)
CDK8/CycC
ULight-CREBtide (7.3)
ULight-GS (5.0)
ULight-rpS6 (4.7)
CDK9/CyclinK
ULight-GS (9.3)
CDK9/CycT1ULight-4E-BP1 (28.1)
ULight-MBP (14.4)
CGK2 (PKG2, PRKG2)ULight-CREBtide (35.8)
ULight-PLK (14.1)
CHK1 (CHEK1)ULight-CREBtide (30.4)
ULight-PKC (15.7)
ULight-Acetyl-CoA Carboxylase (8.1)
CHK2 (CHEK2)ULight-CREBtide (31.8)
ULight-PLK (7.3)
ULight-rpS6 (6.4)
CK1a (CSNK1A1)ULight-Topo IIa (13.7)
CK1-alpha1ULight-Topo IIa (15.0)
CK1d (CSNK1D)ULight-Topo IIa (23.5)
ULight-GS (12.5)
CK1e (CSNK1E)ULight-Topo IIa (23.7)
CK1g1 (CSNK1G1)ULight-Histone H3 (Thr3; 14.6)
CK1g2 (CSNK1G2)ULight-Histone H3 (Thr3; 10.1)
ULight-Topo IIa (7.2)
CK1g3 (CSNK1G3)ULight-Histone H3 (Thr3; 12.9)
CK2-alpha1ULight-Topo IIa (22.0)
CK2a1/b (CSNK2A1/B)ULight-IkBa (28.7)
ULight-Topo IIa (13.8)
ULight-Histone H3 (Thr3; 10.9)
CK2-alpha2ULight-Topo IIa (21.4)
CK2a2/b (CSNK2A2/B)ULight-IkBa (29.6)
ULight-Topo IIa (14.6)
ULight-Histone H3 (Thr3; 10.1)
CLK1ULight-MBP (41.2)
ULight-4E-BP1 (25.7)
ULight-GS (20.1)
CLK2ULight-CREBtide (19.1)
ULight-MBP (11.9)
ULight-Histone H3 (Ser10; 8.3) 2
Kinase Supplier
Supplier Cat. No.
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Carna Biosciences
ProQinase
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Carna Biosciences
ProQinase
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ProQinase
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Invitrogen
Carna Biosciences
Carna Biosciences
Carna Biosciences
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Carna Biosciences
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ProQinase
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Carna Biosciences
02-108
05-107
05-108
05-108
04-102
04-112
0382-0000-1
01-108
05-109
0134-0054-1
0134-0135-1
0134-0055-1
04-103
04-165
04-104
0142-0143-1
04-105
04-106
0356-0389-1
0356-0355-1
0051-0143-1
04-107
04-108
0376-0390-1
PV4335
04-110
01-143
02-117
02-162
03-101
0406-0000-1
03-103
03-104
03-105
03-106
03-107
0124-0000-1
05-184
0412-0000-1
05-185
04-126
04-127
Newly added Kinases highlighted in Green
4
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Kinase ULight Substrate 1
ULight Substrate 2 ULight Substrate 3
CLK3ULight-CREBtide (26.2)
ULight-GS (14.1)
ULight-rpS6 (8.5)
COT (MAP3K8)ULight-p70S6K (Mn2+; 11.0)
ULight-Acetyl-CoA Carboxylase (Mn2+; 7.8) GST-MAP2K1 / ULight-anti-GST (Mn2+; 7.8)
COT GST-MAP2K1 / ULight-anti-GST (Mn2+; 8.0) ULight-p70S6K (Mn2+; 5.3)
ULight-Acetyl-CoA Carboxylase (Mn2+; 4.3)
CRIK (CIT)ULight-Histone H3 (Ser10; 8.3) DAPK1ULight-CREBtide (23.7)
ULight-PLK (18.1)
ULight-Histone H3 (Ser10; 9.3) DAPK1ULight-CREBtide (10.5)
ULight-Histone H3 (Thr3; 9.8)
DAPK2ULight-CREBtide (6.7)
ULight-PLK (6.4)
DAPK3 ULight-CREBtide (13.8)
ULight-Histone H3 (Thr3; 6.6)
ULight-Histone H3 (Ser10; 6.3)
DCAMKL1ULight-CREBtide (34.8)
DCAMKL2ULight-CREBtide (31.4)
DLK (MAP3K12)ULight-CREBtide (36.4)
ULight-PKC (11.6)
ULight-Histone H3 (Thr3; 10.9)
DNA PK3ULight-mTOR (40 U; S/B +/- DNA: 6.6)
DRAK1 (STK17A)ULight-CREBtide (9.2)
ULight-Histone H3 (Thr3; 6.2)
DYRK1AULight-GS (20.1)
ULight-MBP (18.5)
ULight-4E-BP1 (7.4)
DYRK1BULight-4E-BP1 (24.6)
ULight-MBP (23.2)
ULight-GS (19.2)
DYRK2ULight-MBP (33.4)
ULight-4E-BP1 (30.0)
ULight-GS (19.9)
DYRK3ULight-4E-BP1 (39.9)
ULight-MBP (22.0)
ULight-GS (18.3)
DYRK4ULight-4E-BP1 (36.3)
ULight-MBP (26.7)
ULight-GS (20.2)
Erk1 (MAPK3)ULight-4E-BP1 (37.9)
GST-ATF-2 / ULight-anti-GST (37.6)
ULight-MBP (26.6)
Erk2 (MAPK1)ULight-4E-BP1 (37.1)
GST-ATF-2 / ULight-anti-GST (35.1)
ULight-MBP (28.0)
ERK2
GST-ATF-2 / ULight-anti-GST (40.0)
ULight-4E-BP1 (31.9)
ULight-MBP (19.4)
Erk5 (MAPK7)ULight-4E-BP1 (22.4)
Erk7 (MAPK15)ULight-4E-BP1 (22.3)
GCN2ULight-CREBtide (13.5)
ULight-Histone H3 (Thr3; 12.3)
GPRK4 (GRK4)ULight-Topo IIa (18.8)
ULight-CREBtide (15.7)
ULight-p70S6K (14.2)
GPRK6 (GRK6)ULight-Histone H3 (Thr3; 10.8)
ULight-Topo IIa (4.3)
GPRK7 (GRK7)ULight-Histone H3 (Thr3; 10.2)
ULight-Topo IIa (5.9)
GRK5ULight-IkBa (26.5)
ULight-Topo IIa (26.3)
ULight-Histone H3 (Thr3; 11.7)
GSK3a (GSK3A)ULight-4E-BP1 (34.5)
ULight-GS (21.5)
GSK3b (GSK3B)ULight-4E-BP1 (35.9)
ULight-GS (21.9)
Haspin (GSG2)ULight-Histone H3 (Thr3; 15.5)
HGK (MAP4K4)ULight-p70S6K (23.8)
HIPK1ULight-4E-BP1 (30.8)
ULight-MBP (22.9)
ULight-GS (20.1)
HIPK2ULight-MBP (37.4)
ULight-GS (17.4)
HIPK3ULight-4E-BP1 (36.9)
ULight-MBP (22.9)
ULight-GS (21.3)
HIPK4ULight-4E-BP1 (37.7)
ULight-MBP (21.2)
ULight-GS (20.5)
HRIULight-CREBtide (13.3)
ULight-PLK (6.2)
ICKULight-MBP (33.4)
ULight-4E-BP1 (24.7)
ULight-GS (12.6)
IKKa (CHUK)ULight-IkBa (22.4)
ULight-rpS6 (17.9)
ULight-Acetyl-CoA Carboxylase (11.8) IKKbULight-p70S6K (30.4)
ULight-IkBa (25.6)
ULight-rpS6 (22.1)
IKK-betaULight-IkBa (25.5)
IKKe (IKBKE)ULight-p70S6K (22.1)
ULight-rpS6 (20.2)
ULight-PLK (15.8)
Kinase Supplier
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Carna Biosciences
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Invitrogen
Carna Biosciences
Carna Biosciences
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Carna Biosciences
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Carna Biosciences
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Invitrogen
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
ProQinase
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Carna Biosciences
Carna Biosciences
ProQinase
Carna Biosciences
04-128
07-301
0180-0000-1
01-104
02-134
0332-0000-1
PV3614
02-136
02-139
02-140
09-111
V7870
02-137
04-130
04-131
04-132
04-133
PV3871
04-142
04-143
0634-0000-7
04-146
04-147
05-153
01-313
01-315
01-116
PV3824
04-140
04-141
05-111
07-137
04-135
04-136
04-137
04-138
0444-0000-1
04-156
05-112
05-084
0258-0000-1
05-114
Newly added Kinases highlighted in Green
5
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Kinase ULight Substrate 1
ULight Substrate 2 ULight Substrate 3
IKK-epsilonULight-rpS6 (22.0)
ULight-p70S6K (15.5)
ULight-PLK (12.5)
IRAK1ULight-p70S6K (28.3)
ULight-Acetyl-CoA Carboxylase (17.7) ULight-Histone H3 (Thr3; 14.4)
IRAK4ULight-p70S6K (Mn2+; 32.4)
ULight-Acetyl-CoA Carboxylase (31.9) IRAK4ULight-p70S6K (Mn2+; 28.1)
ULight-Acetyl-CoA Carboxylase (23.9) JNK1 (MAPK8)ULight-4E-BP1 (34.5)
GST-ATF-2 / ULight-anti-GST (32.0)
ULight-MBP (21.4)
JNK2 (MAPK9)ULight-4E-BP1 (35.0)
GST-ATF-2 / ULight-anti-GST (32.8)
ULight-MBP (20.8)
JNK3 (MAPK10)
GST-ATF-2 / ULight-anti-GST (33.7)
ULight-4E-BP1 (32.8)
ULight-MBP (19.8)
LATS2ULight-CREBtide (14.8)
ULight-mTOR (5.0)
LRRK2ULight-CREBtide (24.2)
ULight-PLK (19.9)
ULight-Histone H3 (Thr3; 10.4)
LRRK2 [G2019S]ULight-Histone H3 (Thr3; 15.4)
LRRK2 [G2019S]ULight-Histone H3 (Thr3; 15.3)
ULight-CREBtide (8.4)
LZKULight-CREBtide (17.9)
ULight-Histone H3 (Thr3; 8.3)
MAP2K3GST-p38a / ULight-anti-GST (19.8)
MAP2K4GST-p38a / ULight-anti-GST (22.1)
MAP2K6 (MKK6)GST-p38a/ ULight-anti-GST (23.1)
MAP3K1ULight-Acetyl-CoA Carboxylase (5.3) MAP3K2
GST-MAP2K1 / ULight-anti-GST (5.9)
MAP3K6ULight-CREBtide (6.5)
MAP3K14 (NIK)ULight-CREBtide (9.3)
NIK (MAP3K14)ULight-CREBtide (8.5)
MAP4K2ULight-rpS6 (18.3)
ULight-CREBtide (16.0)
ULight-p70S6K (9.2)
MAP4K5 (KHS1)ULight-p70S6K (35.6)
ULight-rpS6 (24.9)
ULight-Acetyl-CoA Carboxylase (23.8) MAPKAPK2ULight-4E-BP1 (32.2)
ULight-MBP (31.4)
ULight-CREBtide (29.1)
MAPKAPK3ULight-CREBtide (27.1)
ULight-rpS6 (13.2)
ULight-PKC (12.3)
MAPKAPK3ULight-CREBtide (26.1)
ULight-mTOR (11.8)
ULight-CDC25 (8.2)
MAPKAPK5ULight-MBP (11.1)
ULight-4E-BP1 (10.1)
MAPKAPK5ULight-Topo IIa (3.6)
ULight-PLK (3.6)
MARK1ULight-PLK (18.1)
ULight-Acetyl-CoA Carboxylase (13.9) ULight-CREBtide (8.9)
MARK1ULight-Acetyl-CoA Carboxylase (9.4) ULight-PLK (9.3)
ULight-CREBtide (8.0)
MARK2 (EMK1)ULight-PLK (18.0)
ULight-Acetyl-CoA Carboxylase (15.6) ULight-CREBtide (8.5)
MARK3 (C-TAK1)ULight-PLK (12.3)
ULight-Acetyl-CoA Carboxylase (11.9) ULight-CDC25 (7.6)
MARK3 (C-TAK1)ULight-Acetyl-CoA Carboxylase (15.1) ULight-PLK (13.2)
ULight-CDC25 (7.1)
MARK4 (MARKL1)ULight-Acetyl-CoA Carboxylase (25.7)
ULight-CREBtide (21.0) ULight-PLK (18.8)
MELKULight-CREBtide (32.9)
MGC42105 (B3KVM1)ULight-CREBtide (25.8)
ULight-PLK (10.8)
ULight-CDC25 (7.4)
MINK (MINK1)ULight-p70S6K (55.1)
ULight-PKC (14.5)
ULight-rpS6 (11.8)
MLK1 (MAP3K9)
GST-MAP2K1 / ULight-anti-GST (11.4)
ULight-rpS6 (5.4)
MLK2 (MAP3K10)ULight-Histone H3 (Thr3; 11.0)
GST-MAP2K1 / ULight-anti-GST (3.2)
MLK3 (MAP3K11)
GST-MAP2K1 / ULight-anti-GST (6.4)
MNK1 (MKNK1)ULight-CREBtide (36.8)
ULight-Histone H3 (Ser10; 9.9) MNK2 (MKNK2)ULight-CREBtide (30.5)
ULight-rpS6 (18.9)
ULight-Histone H3 (Ser10; 13.2)
MOSULight-Topo IIa (9.1)
ULight-CREBtide (6.6)
GST-MAP2K1 / ULight-anti-GST (6.3)
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ProQinase
Carna Biosciences
Carna Biosciences
ProQinase
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Invitrogen
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
ProQinase
Carna Biosciences
Invitrogen
Carna Biosciences
Carna Biosciences
ProQinase
Carna Biosciences
ProQinase
Carna Biosciences
ProQinase
Carna Biosciences
Carna Biosciences
ProQinase
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
0320-0000-1
09-101
09-145
0586-0000-1
04-163
04-164
04-150
01-124
09-110
PV4881
09-146
09-114
07-048
07-044
07-046
07-103
07-004
07-108
07-102
0344-0000-1
07-111
PV3682
02-142
02-143
0633-0000-1
02-144
0380-0000-1
02-120
0432-0000-1
02-121
02-122
0433-0000-1
02-123
02-124
02-125
07-139
09-015
09-116
09-017
02-145
02-146
05-118
Newly added Kinases highlighted in Green
6
www.perkinelmer.com/lance
Kinase ULight Substrate 1
ULight Substrate 2 ULight Substrate 3
MRCKa (CDC42BPA)ULight-Histone H3 (Ser10; 7.5) ULight-CREBtide (6.7)
MSK1 (RPS6KA5)ULight-CREBtide (28.5)
ULight-rpS6 (26.4)
ULight-PLK (20.6)
MSK2 (RPS6KA4)ULight-CREBtide (31.1)
ULight-rpS6 (25.0)
ULight-PLK (19.8)
MST1 (STK4)ULight-p70S6K (13.3)
ULight-PKC (12.8)
ULight-Histone H3 (Thr3; 8.9)
MST1 (STK4)ULight-PKC (11.9)
ULight-Histone H3 (Thr3; 8.4)
ULight-p70S6K (6.5)
MST1 (STK4)ULight-PKC (14.0)
ULight-Histone H3 (Thr3; 6.5)
ULight-rpS6 (5.5)
MST2 (STK3)ULight-Histone H3 (Thr3; 8.0)
ULight-PKC (5.2)
ULight-p70S6K (4.4)
MST2 (STK3)ULight-p70S6K (12.5)
ULight-PKC (9.0)
ULight-Histone H3 (Thr3; 6.2)
MST3 (STK24)ULight-CREBtide (14.7)
ULight-PKC (9.3)
ULight-rpS6 (8.9)
MST4 (MASK)ULight-PKC (14.6)
ULight-rpS6 (8.5)
mTOR (1362-end)ULight-4E-BP1 (Mn2+; 36.9)
ULight-p70S6K (Mn2+; 18.3)
MYT1 (PKMYT1)ULight-CREBtide (10.6)
NDR1 (STK38)ULight-CREBtide (10.5)
ULight-Histone H3 (Ser10; 8.7) NDR2ULight-Histone H3 (Ser10; 8.0)
ULight-CREBtide (7.0)
NEK1ULight-p70S6K (60.0)
ULight-Acetyl-CoA Carboxylase (24.0) ULight-rpS6 (18.6)
NEK2ULight-p70S6K (43.3)
ULight-CREBtide (31.2)
ULight-rpS6 (18.2)
NEK3ULight-p70S6K (26.9)
ULight-4E-BP1 (18.2)
NEK4ULight-p70S6K (40.1)
NEK6ULight-p70S6K (32.0)
NEK7ULight-p70S6K (33.4)
NEK9ULight-p70S6K (48.9)
ULight-rpS6 (16.5)
ULight-Acetyl-CoA Carboxylase (13.0)
NEK11ULight-p70S6K (20.0)
ULight-Histone H3 (Thr3; 14.9)
NLKULight-CREBtide (9.4)
ULight-PLK (1.3)
NLKULight-CREBtide (15.4)
ULight-PLK (6.0)
Nuak2 (SNARK)ULight-CREBtide (23.9)
ULight-Histone H3 (Thr3; 7.2)
SNARK (Nuak2)ULight-CREBtide (14.8)
ULight-Histone H3 (Thr3; 5.8)
ULight-PLK (5.5)
OSR1 (OXSR1)ULight-Histone H3 (Thr3; 5.6)
p38a (MAPK14)ULight-4E-BP1 (37.0)
GST-ATF-2 / ULight-anti-GST (32.1)
ULight-MBP (27.9)
p38-alpha (MAPK14)
GST-ATF-2 / ULight-anti-GST (36.7)
ULight-4E-BP1 (33.1)
ULight-MBP (5.4)
p38b (MAPK11)ULight-MBP (40.6)
GST-ATF-2 / ULight-anti-GST (36.9)
ULight-4E-BP1 (32.6)
p38-beta (MAPK11)
GST-ATF-2 / ULight-anti-GST (36.8)
ULight-4E-BP1 (35.8)
ULight-MBP (14.2)
p38d (MAPK13)ULight-MBP (41.2)
GST-ATF-2 / ULight-anti-GST (35.1)
ULight-4E-BP1 (30.7)
p38g (MAPK12)ULight-4E-BP1 (37.7)
GST-ATF-2 / ULight-anti-GST (37.0)
ULight-MBP (33.6)
p70S6K (S6K; RPS6KB1)ULight-CREBtide (30.2)
ULight-rpS6 (18.1)
ULight-Histone H3 (Ser10; 9.4)
p70S6Kb (RPS6KB2)ULight-CREBtide (28.6)
ULight-rpS6 (23.0)
ULight-Crosstide (13.9)
PAK1ULight-PLK (12.3)
ULight-Histone H3 (Ser10; 4.5) PAK1ULight-PLK (15.6)
PAK2ULight-PLK (16.7)
ULight-Histone H3 (Ser10; 4.9)
PAK3ULight-PLK (19.9)
ULight-Histone H3 (Ser10; 12.2) ULight-GS (9.2)
PAK4ULight-CREBtide (12.2)
ULight-PLK (4.3)
PAK5 (PAK7)ULight-PLK (18.5)
PAK61ULight-PLK (5h; 7.0)
Kinase Supplier
Supplier Cat. No.
Carna Biosciences
Carna Biosciences
Carna Biosciences
Millipore
Carna Biosciences
ProQinase
Carna Biosciences
ProQinase
Carna Biosciences
Carna Biosciences
Millipore
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
ProQinase
Carna Biosciences
ProQinase
Carna Biosciences
Carna Biosciences
ProQinase
Carna Biosciences
ProQinase
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
ProQinase
Carna Biosciences
Carna Biosciences
ProQinase
Carna Biosciences
Carna Biosciences
01-107
01-147
01-148
14-624
07-116
0667-0000-1
07-117
0699-0000-1
07-118
07-119
14-770
05-176
01-125
01-126
05-123
05-226
05-127
05-128
05-130
05-131
05-133
05-125
04-151
0253-0000-1
02-127
0414-0000-1
07-122
04-152
0443-0000-3
04-153
0455-0000-3
04-154
04-155
01-154
01-155
07-123
0357-0000-1
07-124
07-125
0365-0000-1
07-127
07-128
Newly added Kinases highlighted in Green
7
www.perkinelmer.com/lance
Kinase ULight Substrate 1
ULight Substrate 2 ULight Substrate 3
PAK6ULight-Histone H3 (Thr3; 8.5)
ULight-PLK (5.5)
PASKULight-CREBtide (31.0)
ULight-Crosstide (12.6)
ULight-mTOR (10.4)
PBKULight-CREBtide (16.3)
PBK1ULight-Acetyl-CoA Carboxylase (5 h; 7.1) PEK (EIF2AK3)ULight-p70S6K (51.9)
ULight-Topo IIa (18.0)
ULight-Acetyl-CoA Carboxylase (15.4) PGK (PRKG1)ULight-CREBtide (37.0)
ULight-PLK (11.1)
PHKG1ULight-CREBtide (31.3)
ULight-Histone H3 (Thr3; 8.5)
ULight-rpS6 (7.0)
PHKG2ULight-CREBtide (31.7)
ULight-rpS6 (15.6)
ULight-mTOR (9.7)
PIM1ULight-CREBtide (34.7)
ULight-mTOR (8.5)
ULight-rpS6 (8.2)
PIM1ULight-CREBtide (31.2)
ULight-rpS6 (21.5)
ULight-mTOR (12.6)
PIM2ULight-CREBtide (30.3)
ULight-rpS6 (20.9)
ULight-mTOR (11.3)
PIM2ULight-CREBtide (14.6)
ULight-rpS6 (10.4)
ULight-mTOR (6.6)
PIM3ULight-CREBtide (31.0)
ULight-rpS6 (24.7)
ULight-mTOR (11.7)
PKACa (PRKACA)ULight-CREBtide (24.3)
ULight-rpS6 (16.8)
ULight-PLK (16.6)
PKACb (PRKACB)ULight-rpS6 (23.7)
ULight-PLK (20.8)
ULight-CREBtide (15.7)
PKCa (PRKCA)ULight-CREBtide (28.1)
ULight-rpS6 (19.4)
ULight-PKC (14.5)
PKCb1 (PRKCB1)ULight-CREBtide (30.5)
ULight-PKC (15.6)
PKCb2 (PRKCB2)ULight-CREBtide (26.1)
ULight-rpS6 (19.1)
ULight-PKC (14.0)
PKC-beta2 (PRKCB2)ULight-CREBtide (31.4)
ULight-PKC (16.2)
ULight-rpS6 (2.9)
PKCd (PRKCD)ULight-CREBtide (29.4)
ULight-PKC (14.8)
PKCe (PRKCE)ULight-CREBtide (31.0)
ULight-PKC (14.9)
PKACgULight-CREBtide (33.4)
ULight-rpS6 (24.2)
ULight-PLK (21.3)
PKCg (PRKCG)ULight-CREBtide (29.7)
ULight-PKC (15.6)
ULight-rpS6 (9.1)
PKC-gamma (PRKCG)ULight-CREBtide (26.1)
ULight-PKC (13.5)
ULight-rpS6 (10.2)
PKCh (PRKCH)ULight-CREBtide (29.5)
ULight-PKC (14.3)
ULight-rpS6 (9.5)
PKC-eta (PRKCH)ULight-rpS6 (22.4)
ULight-PLK (13.2)
ULight-PKC (10.6)
PKCi (PRKCI)ULight-CREBtide (29.4)
ULight-PKC (15.4)
ULight-rpS6 (8.9)
PKCu (PRKCQ)ULight-CREBtide (30.4)
ULight-PKC (15.1)
ULight-rpS6 (12.3)
PKCz (PRKCZ)ULight-CREBtide (22.4)
ULight-PKC (14.1)
ULight-rpS6 (12.9)
PKC-zetaULight-CREBtide (18.6)
ULight-PKC (14.1)
PKC-mu (PKD-1, PRKD1)ULight-CREBtide (23.2)
ULight-PLK (8.4)
ULight-PKC (5.6)
PKD1 (PRKD1, PKC-mu)ULight-CREBtide (21.6)
ULight-PLK (14.4)
PKC-nu (PRKD3, EPK2)ULight-CREBtide (23.1)
ULight-PLK (5.4)
PKD3 (PRKD3, PKC-nu)ULight-CREBtide (31.4)
ULight-Histone H3 (Thr3; 8.8)
ULight-PLK (6.0)
PKD2 (PRKD2)ULight-PLK (10.3)
ULight-CREBtide (9.4)
PKN1 (PRK1)ULight-CREBtide (32.6)
ULight-PKC (10.5)
ULight-rpS6 (7.5)
PRK1 (PKN1)ULight-CREBtide (31.8)
ULight-rpS6 (17.9)
ULight-PKC (14.0)
PKN2ULight-CREBtide (32.3)
PKN3ULight-CREBtide (33.5)
ULight-rpS6 (20.0)
ULight-PKC (15.4)
PKR (EIF2AK2)ULight-Histone H3 (Thr3; 10.8)
ULight-CREBtide (6.7)
PLK1ULight-p70S6K (24.8)
ULight-Topo IIa (23.0)
ULight-Histone H3 (Thr3; 13.4)
PLK1ULight-Topo IIa (7.0)
ULight-Histone H3 (Thr3; 5.7)
Kinase Supplier
Supplier Cat. No.
ProQinase
Carna Biosciences
ProQinase
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
ProQinase
Carna Biosciences
ProQinase
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
ProQinase
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
ProQinase
Carna Biosciences
ProQinase
Carna Biosciences
Carna Biosciences
Carna Biosciences
ProQinase
ProQinase
Carna Biosciences
ProQinase
Carna Biosciences
Carna Biosciences
Carna Biosciences
ProQinase
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
ProQinase
0423-0000-1
02-128
0185-0000-1
05-168
05-155
01-142
02-152
02-153
02-054
0186-0000-1
02-155
0223-0000-1
02-156
01-127
01-128
01-133
01-134
01-165
0116-0000-1
01-135
01-136
01-129
01-137
0204-0000-1
01-138)
0420-0000-1
01-139
01-140
01-141
0117-0000-1
0115-0000-1
02-157
0661-0000-1
02-159
02-158
01-144
0207-0000-1
01-145
01-146
05-156
05-157
0183-0000-1
Newly added Kinases highlighted in Green
8
www.perkinelmer.com/lance
Kinase ULight Substrate 1
ULight Substrate 2 ULight Substrate 3
PLK2 (SNK)ULight-Topo IIa (22.9)
ULight-CREBtide (11.6)
PLK3ULight-IkBa (27.5)
ULight-Topo IIa (20.4)
ULight-4E-BP1 (15.0)
PLK3ULight-Topo IIa (18.6)
ULight-Histone H3 (Thr3; 6.1)
ULight-4E-BP1 (5.1)
PLK4 (SAK)ULight-CREBtide (10.4)
ULight-PLK (1.9)
SAK (PLK4) ULight-CREBtide (11.4)
ULight-PLK (5.8)
PRKXULight-CREBtide (34.1)
ULight-PLK (19.7)
ULight-PKC (13.7)
QIK (SNF1LK2) ULight-CREBtide (29.6)
ULight-Acetyl-CoA Carboxylase (28.8)
ULight-PLK (14.1)
SNF1LK2 (QIK)ULight-CREBtide (32.2)
ULight-Acetyl-CoA Carboxylase (24.8) ULight-PLK (14.5)
RAF1
GST-MAP2K1/ ULight-anti-GST (9.3)
RIPK2ULight-CREBtide (8.4)
RIPK5ULight-CREBtide (9.7)
ULight-p70S6K (5.7)
ROCK1ULight-CREBtide (34.8)
ULight-rpS6 (23.4)
ULight-PLK (19.6)
ROCK2ULight-CREBtide (32.7)
ULight-rpS6 (20.0)
ULight-PLK (16.9)
RSK1 (RPS6KA1)ULight-CREBtide (32.1)
ULight-rpS6 (22.0)
ULight-PLK (20.5)
RSK2 (RPS6KA3)ULight-CREBtide (30.0)
ULight-rpS6 (20.9)
ULight-Crosstide (18.8)
RSK3 (RPS6KA2)ULight-CREBtide (32.0)
ULight-rpS6 (25.9)
ULight-PLK (18.5)
RSK4 (RPS6KA6)ULight-CREBtide (32.0)
ULight-rpS6 (23.8)
ULight-PLK (18.4)
SGKULight-CREBtide (32.7)
ULight-rpS6 (20.8)
ULight-PLK (20.6)
SGK2ULight-CREBtide (32.5)
ULight-PLK (21.3)
ULight-rpS6 (21.1)
SGK3 (SGKL)ULight-CREBtide (28.0)
ULight-rpS6 (22.7)
ULight-PLK (17.7)
SIKULight-CREBtide (17.8)
ULight-Acetyl-CoA Carboxylase (9.2)
ULight-PLK (7.4)
SLKULight-CREBtide (31.0)
skMLCK (MYLK)2ULight-CREBtide (35.8)
SRPK1ULight-CREBtide (20.0)
SRPK2ULight-CREBtide (14.3)
STK33ULight-rpS6 (13.1)
ULight-mTOR (9.5)
ULight-CREBtide (8.0)
STK33ULight-CREBtide (8.3)
TAK1-TAB1 (MAP3K7)ULight-p70S6K (5.6)
TAOK2ULight-p70S6K (15.6)
ULight-CREBtide (9.0)
ULight-PKC (5.7)
TAOK3ULight-CREBtide (21.2)
ULight-Histone H3 (Thr3; 9.3)
ULight-p70S6K (7.9)
TBK1ULight-p70S6K (34.5)
ULight-rpS6 (24.8)
ULight-Acetyl-CoA Carboxylase (24.5) TGFßR1 (TGFBR1)ULight-Topo IIa (21.3)
TGFßR2 (TGFBR2)ULight-CREBtide (6.7)
TLK1ULight-Topo IIa (17.4)
ULight-Histone H3 (Thr3; 15.2)
ULight-p70S6K (9.4)
TLK2ULight-Topo IIa (17.9)
ULight-Histone H3 (Thr3; 17.2)
ULight-p70S6K (13.8)
TSK2 (TSSK2)ULight-CREBtide (32.1)
ULight-PLK (20.2)
ULight-Acetyl-CoA Carboxylase (15.6) TSSK2 (TSK2)ULight-CREBtide (31.0)
ULight-Acetyl-CoA Carboxylase (17.9) ULight-PLK (16.2)
TSSK1ULight-CREBtide (20.4)
ULight-PLK (17.9)
ULight-Acetyl-CoA Carboxylase (16.2) TSSK3ULight-CREBtide (32.5)
ULight-Acetyl-CoA Carboxylase (27.3)
ULight-PLK (20.0)
TTBK1ULight-Topo IIa (24.9)
ULight-Histone H3 (Thr3; 17.8)
ULight-p70S6K (13.4)
TTBK2ULight-Topo IIa (24.2)
ULight-Histone H3 (Thr3; 17.4)
ULight-rpS6 (10.3)
ULK3ULight-Histone H3 (Thr3; 6.5)
Kinase Supplier
Supplier Cat. No.
Carna Biosciences
05-158
Carna Biosciences
05-159
ProQinase
0437-0000-1
Carna Biosciences
05-160
ProQinase
0306-0000-1
Carna Biosciences
01-130
Carna Biosciences
02-129
Invitrogen
PV4792
Carna Biosciences
09-125
Carna Biosciences
09-128
Invitrogen
PV4678
Carna Biosciences
01-109
Carna Biosciences
01-110
Carna Biosciences
01-149
Carna Biosciences
01-150
Carna Biosciences
01-151
Carna Biosciences
01-152
Carna Biosciences
01-158
Carna Biosciences
01-159
Carna Biosciences
01-160
Carna Biosciences
02-131
Carna Biosciences
07-129
Carna Biosciences
02-150
ProQinase
0434-0000-1
ProQinase
0381-0000-1
Millipore14-671
Carna Biosciences
02-301
Carna Biosciences
09-019
Carna Biosciences
07-133
Carna Biosciences
07-135
Carna Biosciences
05-115
Carna Biosciences
09-141
Carna Biosciences
09-142
Carna Biosciences
05-166
Carna Biosciences
05-167
Carna Biosciences
02-165
ProQinase
0220-0000-1
Carna Biosciences
02-364
Carna Biosciences
02-166
Carna Biosciences
03-108
Carna Biosciences
03-109
Carna Biosciences
05-173
Newly added Kinases highlighted in Green
9
www.perkinelmer.com/lance
Kinase ULight Substrate 1
ULight Substrate 2 ULight Substrate 3
VRK1ULight-CREBtide (17.0)
ULight-Histone H3 (Thr3; 8.7)
VRK2ULight-Histone H3 (Thr3; 15.1)
WEEULight-CREBtide (10.0)
WNK1ULight-CREBtide (7.4)
WNK2ULight-CREBtide (21.4)
WNK3ULight-CREBtide (4.8)
WNK4ULight-CREBtide (10.3)
YSK1 (STK25)ULight-CREBtide (21.8)
ULight-rpS6 (14.3)
ULight-p70S6K (5.7)
ZAKULight-p70S6K (36.1)
ULight-Histone H3 (Thr3; 15.5)
ULight-mTOR (11.4)
Kinase Supplier
Supplier Cat. No.
ProQinase
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
Carna Biosciences
0302-0000-1
03-111
05-177
05-179
05-180
05-181
05-182
07-136
09-120
Kinase reaction time is 5h
CaCl2 and Calmodulin at 1 mM and 0.5 µM, respectively, are added to the kinase reactions
3
DNA PK: assay buffer and conditions as recommended by Promega. ATP: 200 μM final, kinase reaction incubation time 1 h
Kinases were assayed in standard conditions, using reagent concentrations and kinase buffer described below. All modifications to standard assay conditions are indicated in the table.
1
2
Kinase assay components:
Kinases: 20 nM (various suppliers)
ATP: 200 µM
ULight substrates in kinase reactions: 50 nM (100 nM for ULight-CDC25)
GST-protein substrates in kinase reactions: 30 nM (when indicated)
ULight-anti-GST antibody in assay: 30 nM (when indicated)
Europium anti-phospho-substrate antibodies in assay: 2 nM
Kinase buffer (pH 7.5):
50 mM HEPES
10 mM MgCl2
3 mM MnCl2 (when indicated)
1 mM EGTA
2 mM DTT and 0.01% Tween 20
After incubation at 23°C for 1 h (or 5 h when indicated), kinase reactions were stopped with EDTA. Antibodies were added and incubated for 1 h prior to detection with the EnVision.
NOTE: All kinases were sourced from commercial vendors from commercially available lots. In many cases, the same kinase was tested from multiple vendors. If the results obtained with kinases from
different vendors were very different in our testing, we have shown all suppliers’ data. If the results were similar, we have shown only one representative set of test results. We encourage you to qualify
your kinase activity to ensure that that the assay signal is specific to the enzyme tested.
Newly added Kinases highlighted in Green
10
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LANCE® Ultra products used for this study:
Labeled Antibody
Description
ULight-CREBtideULight-CREBtide (Ser133)
Europium-anti-phospho-CREB
Europium-anti-phospho-CREB (Ser133)
ULight-MBPULight-Myelin Basic Protein Peptide
Europium-anti-phospho-MBP
Europium-anti-phospho-Myelin Basic Protein
ULight-CrosstideULight-Crosstide (GSK-3a Ser21)
Europium-anti-phospho-Crosstide
Europium-anti-phospho-Crosstide (GSK-3a Ser21)
ULight-PLKULight-PLK (Ser137) Peptide
Europium-anti-phospho-PLK
Europium-anti-phospho-PLK (Ser137)
ULight-PKCULight-PKC Substrate (Ala25Ser)
Europium anti-phospho-PKC
Europium anti-phospho-PKC Peptide (Ala25Ser)
ULight-IkBaULight-IkappaB-alpha (Ser32/36) Peptide
Europium-anti-phospho-IkBa
Europium-anti-phospho-IkappaB-alpha (Ser32/36)
ULight-Histone H3 (Thr3/Ser10)
ULight-Histone H3 (Thr3/Ser10) Peptide
Europium-anti-phospho-Histone H3 (Ser10)
Europium-anti-phospho-Histone H3 (Ser10)
Europium-anti-phospho-Histone H3 (Thr3)
Europium-anti-phospho-Histone H3 (Thr3
ULight-mTORULight-mTOR (Ser2448) Peptide
Europium-anti phospho-mTOR
Europium-anti phospho-mTOR (Ser2448)
ULight-p70S6KULight-p70S6K (Thr389) Peptide
Europium-anti-phospho-p70S6K
Europium-anti-phospho-p70S6K (Thr389)
ULight-Acetyl-CoA Carboxylase
ULight-Acetyl-CoA Carboxylase (Ser79) SAMS Peptide
Europium-anti-phospho-Acetyl-CoA Carboxylase
Europium-anti-phospho-Acetyl-CoA Carboxylase (Ser79)
ULight-Cdc25CULight-Cdc25C (Ser216) Peptide
Europium-anti-phospho-(Ser) 14-3-3 binding motif
Europium-anti-phospho-(Ser) 14-3-3 binding motif, mAb 4E2
ULight-4E-BP1ULight-eIF4E-binding protein 1 (Thr37/46) Peptide
Europium-anti-phospho-4E-BP1
Europium-anti-phospho-eIF4E-binding protein 1 (Thr37/46)
ULight-rpS6ULight-40S Ribosomal Protein S6 (pSer235/Ser236) Peptide
Europium-anti-phospho-rpS6
Europium-anti-phospho-40S Ribosomal Protein S6 (Ser235/236)
ULight-Topo IIaULight-DNA Topoisomerase 2-alpha (Thr1342) Peptide
Europium-anti-phospho-Topo IIa
Europium-anti-phospho-DNA Topoisomerase 2-alpha (Thr1342)
ULight-anti-GSTULight-anti-GST
Europium-anti-phospho-ATF-2
Europium-anti-phospho-ATF-2 (Thr53)
ATF-2 Fusion Protein
Europium-anti-phospho-p38a
Europium-anti-phospho-MAP Kinase p38-alpha (Thr180/Tyr182)
GST-p38a (MAPK14), inactive
Europium-anti-phospho-MEK1/2
Europium-anti-phospho-MEK1/2 (Ser217/221)
GST-MAP2K1, inactive
ULight-Glycogen Synthase
ULight-Glycogen Synthase (Ser641/pSer657) Peptide
Europium anti-phospho-Glycogen Synthase
Europium anti-phospho-Glycogen Synthase (Ser641)
11
Catalog No. Phospho-Motif
TRF0107
TRF0200
TRF0109
TRF0201
TRF0106
TRF0202
TRF0110
TRF0203
TRF0108
TRF0207
TRF0113
TRF0206
TRF0125
TRF0210
TRF0211
TRF0119
TRF0209
TRF0126
TRF0214
TRF0118
TRF0208
TRF0123
AD0192
TRF0128
TRF0216
TRF0129
TRF0217
TRF0130
TRF0218
TRF0104
TRF0212
Cell Signaling Technology cat. 9224
TRF0219
Carna BioSciences cat. 04-152-10
TRF0213
Carna BioSciences cat. 07-141-10
TRF0131
TRF0220
RRPSYRK
VTPRTPPP
PRTSSFAE
RRRSLLE
RKGSLRQ
RHDSGLDSM
ARTKQTARKSTG
TRTDSYSAG
FLGFTYVAP
RSAMSGLHL
YRSPSMPEN
(R/K)XXSXP
DYSTTPGGTLFSTTPGTRI
KRRRLpSSSLRAS
FDEKTDDEDF
PASVPPSPSLSRHSSPHQ(pS)ED
www.perkinelmer.com/lance
LANCE Ultra Tyrosine Kinase Selection Guide
How to use this guide:
1.Locate your kinase
• In many cases, up to three commercial kinase vendors have been tested.
• Many common aliases are shown in parenthesis.
2.Best performing ULight ™ substrates are listed for each enzyme according
to performance criteria where signal to background (S/B) ratios are greater
than 3.0
• S/B ratios (Signal at 665 nm / minus ATP control at 665 nm) are indicated
in parenthesis.
• All S/B ratios were obtained at fixed experimental conditions unless
otherwise noted (see page 16).
If you cannot find your kinase of interest, please ask your PerkinElmer sales
specialist, as our list continues to expand. Two kits are available for
testing purposes:
• KinaSelect Ser/Thr kit (5 x 250 data points, TRF0300-C)
– 5 ULight-labeled Ser/Thr kinase specific substrates + 5 matching
Europium-labeled anti-phospho antibodies
• KinaSelect TK kit (1,000 data points, TRF0301-D)
– 1 ULight-labeled kinase specific substrate + 1 matching
Europium-labeled anti-phospho antibody
3.Based on your substrate choice, find the corresponding Europium-labeled
anti-phospho antibody on page 16 (LANCE Ultra products used for
this study) to identify the TR-FRET donor anti-phospho antibody for your
LANCE Ultra assay.
12
www.perkinelmer.com/lance
LANCE® Ultra Reagents for Tyr Kinase Assays
Up to three kinase substrates are listed based on signal to background (S/B) ratios obtained with commercially available enzymes.
S/B ratios (Signal at 665 nm / minus ATP control at 665 nm) are indicated in parentheses.
ULight Substrate 2
ULight Substrate 3
ABL [0.25]ULight-poly GT (13.1)
ULight-poly GAT (11.1)
ULight-TK (4.1)
Carna Biosciences
08-001
ABL (T315I, ABL1) [0.5]ULight-poly GT (6.6)
ULight-poly GAT (6.4)
ULight-TK (4.8)
Carna Biosciences
08-093
ACK [20]ULight-poly GT (27.3)
ULight-poly GAT (27.2)
ULight-TK (17.3)
Carna Biosciences
08-103
ALK [4]ULight-IRS-1 (16.8)
ULight-poly GT (9.8)
ULight-TK (8.8)
Carna Biosciences
08-105
ARG [0.5]ULight-poly GT (13.2)
ULight-poly GAT (12.7)
ULight-TK (10.6)
Carna Biosciences
08-102
AXL [4]ULight-poly GT (9.8)
ULight-poly GAT (5.6)
Carna Biosciences
08-107
BLK [0.5]ULight-TK (31.0)
ULight-poly GT (16.5)
ULight-CDK1 (9.6)
Carna Biosciences
08-164
BMX [0.5]ULight-poly GT (13.2)
ULight-poly GAT (9.0)
ULight-TK (7.1)
Carna Biosciences
08-179
BRK [3]ULight-poly GT (17.4)
ULight-poly GAT (3.0)
Carna Biosciences
08-165
BTK [0.25]ULight-poly GAT (19.6)
ULight-poly GT (11.8)
ULight-TK (6.8)
Carna Biosciences
08-080
CSK [20]ULight-TK (27.2)
ULight-poly GAT (20.3)
ULight-poly GT (10.4)
Carna Biosciences
08-111
CTK [20]ULight-poly GAT (18.8)
Kinase [Concentration, nM]
ULight Substrate 1
Kinase Supplier
Supplier Cat. No.
ULight-poly GT (12.6)
Carna Biosciences
08-012
DDR1 [0.5]ULight-poly GAT (3.0)
Carna Biosciences
08-113
DDR2 [0.5]ULight-poly GAT (5.1)
ULight-IRS-1 (4.9)
ULight-poly GT (4.1)
Carna Biosciences
08-114
EGFR [0.5]ULight-JAK-1 (19.0)
ULight-TK (10.0)
ULight-poly GAT (5.0)
Carna Biosciences
08-115
EGFR (T790M) [0.5]ULight-poly GT (18.1)
ULight-poly GAT (10.8)
ULight-JAK-1 (3.9)
Carna Biosciences
08-194
EphA1 [0.5]ULight-poly GT (24.1)
ULight-poly GAT (11.4)
ULight-TK (10.8)
Carna Biosciences
08-120
EphA2 [0.25]ULight-TK (26.9)
ULight-poly GT (12.3)
ULight-poly GAT (7.1)
Carna Biosciences
08-121
EphA3 [0.5]ULight-TK (22.4)
ULight-poly GT (17.4)
ULight-poly GAT (8.0)
Carna Biosciences
08-122
EphA4 [0.5]ULight-TK (31.5)
ULight-poly GT (12.6)
ULight-poly GAT (11.3)
Carna Biosciences
08-123
EphA5 [1]ULight-TK (35.5)
ULight-poly GAT (21.9)
ULight-JAK-1 (21.2)
Carna Biosciences
08-124
EphA6 [0.5]ULight-poly GAT (15.5)
ULight-poly GT (14.2)
ULight-TK (5.4)
Carna Biosciences
08-125
EphA7 [0.5]ULight-poly GT (22.7)
ULight-TK (17.2)
ULight-poly GAT (11.1)
Carna Biosciences
08-126
EphA8 [0.25]ULight-TK (32.3)
ULight-poly GT (16.4)
ULight-poly GAT (7.8)
Carna Biosciences
08-127
EphB1 [0.5]ULight-TK (29.9)
ULight-poly GT (25.1)
ULight-CDK1 (14.1)
Carna Biosciences
08-128
EphB2 [0.25]ULight-TK (28.8)
ULight-poly GT (27.9)
ULight-poly GAT (13.2)
Carna Biosciences
08-129
EphB3 [0.1]ULight-poly GT (29.0)
ULight-TK (24.7)
ULight-poly GAT (11.2)
Carna Biosciences
08-130
EphB4 [0.5]ULight-TK (29.8)
ULight-poly GT (13.7)
ULight-CDK1 (12.6)
Carna Biosciences
08-131
FAK [3]ULight-poly GT (20.0)
ULight-poly GAT (13.0)
Carna Biosciences
08-037
FER [0.25]ULight-TK (31)
ULight-poly GAT (17.1)
ULight-CDK1 (16.1)
Carna Biosciences
08-139
FES [1]ULight-TK (29.4)
ULight-poly GAT (19.0)
ULight-poly GT (17.1)
Carna Biosciences
08-140
13
www.perkinelmer.com/lance
Kinase [Concentration, nM]ULight Substrate 1
ULight Substrate 2
ULight Substrate 3
Kinase Supplier
Supplier Cat. No.
FGFR1 [0.5]ULight-JAK-1 (30.8)
ULight-TK (26.4)
ULight-IRS-1 (13.5)
Carna Biosciences
08-133
FGFR2 [0.25]ULight-JAK-1 (33.2)
ULight-TK (27.1)
ULight-IRS-1 (15.5)
Carna Biosciences
08-134
FGFR3 [0.25]ULight-JAK-1 (27.6)
ULight-TK (24.4)
ULight-poly GT (16.0)
Carna Biosciences
08-135
FGFR4 [3]ULight-JAK-1 (20.8)
ULight-TK (17.7)
ULight-poly GT (9.2)
Carna Biosciences
FGR [0.1]ULight-TK (29.3)
ULight-poly GT (25.1)
ULight-poly GAT (17.7)
Carna Biosciences
08-166
FLT1 [0.25]ULight-poly GT (22.2)
ULight-poly GAT (15.4)
ULight-JAK-1 (10.0)
Carna Biosciences
08-189
FLT3 [0.1]ULight-TK (25.9)
ULight-JAK-1 (25.0)
ULight-poly GT (20.8)
Carna Biosciences
08-154
FLT4 [0.25]ULight-JAK-1 (36.9)
ULight-TK (20.0)
ULight-IRS-1 (11.4)
Carna Biosciences
08-190
FMS (CSFR) [0.25]ULight-poly GT (14.6)
ULight-JAK-1 (6.3)
ULight-poly GAT (6.2)
Carna Biosciences
08-155
FRK [0.5]ULight-poly GAT (22.9)
ULight-poly GT (20.8)
ULight-TK (9.7)
Carna Biosciences
08-167
FYN [0.25]ULight-TK (25.9)
ULight-CDK1 (19.7)
ULight-poly GT (12.8)
Carna Biosciences
08-068
HCK [0.25]ULight-TK (29.7)
ULight-poly GT (21.3)
ULight-poly GAT (13.2)
Carna Biosciences
08-169
HER2 [10]ULight-poly GT (9.3)
ULight-poly GAT (8.4)
Carna Biosciences
08-166
HER4 (ERBB4) [0.1]ULight-poly GT (25.2)
ULight-poly GAT (14.4)
ULight-TK (3.1)
Carna Biosciences
08-118
IGF1R [0.25]ULight-TK (25.1)
ULight-poly GT (21.8)
ULight-IRS-1 (15.2)
Carna Biosciences
08-141
INSR [20]ULight-poly GAT (25.4)
ULight-poly GT (18.3)
ULight-TK (4.0)
Carna Biosciences
08-242
IRR [0.5]ULight-poly GT (21.6)
ULight-TK (19.2)
ULight-IRS-1 (15.6)
Carna Biosciences
08-143
ITK [0.5]ULight-poly GT (9.8)
ULight-poly GAT (8.5)
ULight-TK (8.4)
Carna Biosciences
08-181
JAK1 [7]ULight-JAK-1 (19.5)
ULight-poly GT (17.6)
ULight-poly GAT (16.4)
Carna Biosciences
08-144
JAK2 [1]ULight-JAK-1 (41.8)
ULight-TK (35.5)
ULight-IRS-1 (21.4)
Carna Biosciences
08-045
JAK3 [0.5]ULight-JAK-1 (37.3)
ULight-TK (16.1)
ULight-poly GAT (6.9)
Carna Biosciences
08-046
KDR [0.25]ULight-JAK-1 (30.8)
ULight-TK (16.9)
ULight-poly GT (8.6)
Carna Biosciences
08-191
KIT [2]ULight-poly GAT (21.7)
ULight-poly GT (13.0)
ULight-JAK-1 (3.3)
Carna Biosciences
08-056
LCK [0.5]ULight-poly GAT (16.5)
ULight-TK (12.3)
ULight-poly GT (10.4)
Carna Biosciences
08-170
LTK [1]ULight-poly GAT (25.1)
ULight-poly GT (21.3)
ULight-TK (13.7)
Carna Biosciences
08-106
LYNa [0.5]ULight-TK (32.5)
ULight-poly GT (15.6)
ULight-CDK1 (14.3)
Carna Biosciences
08-171
LYNb [1]ULight-TK (33.1)
ULight-CDK1 (24.0)
ULight-poly GT (18.6)
Carna Biosciences
08-172
MATK (Hyl) [1]ULight-poly GT (20.7) ULight-poly GAT (8.8) `
Invitrogen
PV3370
MER [0.25]ULight-poly GT (29.2)
ULight-poly GAT (15.9)
ULight-TK (12.9)
Carna Biosciences
08-108
MET [0.5]ULight-TK (24.2)
ULight-JAK-1 (19.7)
ULight-poly GT (17.0)
Carna Biosciences
08-151
MUSK [1]ULight-poly GAT (10.6)
ULight-JAK-1 (3.9)
ULight-TK (3.1)
Carna Biosciences
08-153
PDGFRa [0.5]ULight-poly GT (17.8)
ULight-poly GAT (10.6)
ULight-JAK-1 (9.0)
Carna Biosciences
08-157
PDGFRb [0.25]ULight-poly GAT (12.5)
ULight-poly GT (11.2)
ULight-TK (4.7)
Carna Biosciences
08-158
PTK2B (FAK2) [0.25] ULight-polyGT (19.2) ULight-poly GAT (12.3)
Invitrogen
PV4567
PYK2 [0.25]ULight-poly GT (15.5)
ULight-poly GAT (7.7)
Carna Biosciences
08-138
14
08-136
www.perkinelmer.com/lance
ULight Substrate 3
Kinase Supplier
Supplier Cat. No.
RET [0.5]ULight-JAK-1 (23.6)
ULight-TK (20.3)
ULight-IRS-1 (6.9)
Carna Biosciences
08-159
RON [0.25]ULight-poly GT (27.7)
ULight-poly GAT (25.7)
ULight-JAK-1 (4.9)
Kinase [Concentration, nM]ULight Substrate 1
ULight Substrate 2
ROR1 [20]ULight-poly GT (3.4)
Carna Biosciences
08-152
Carna Biosciences
08-160
ROR2 [20]ULight-poly GT (4.2)
Carna Biosciences
08-161
ROS [0.25]ULight-TK (24.5)
ULight-poly GT (23.8)
ULight-IRS-1 (20.5)
Carna Biosciences
08-163
SRC [0.5]ULight-TK (31.7)
ULight-poly GAT (18.4)
ULight-CDK1 (16.6)
Carna Biosciences
08-173
SRM [0.5]ULight-TK (32.5)
ULight-poly GAT (20.1)
ULight-poly GT (18.3)
Carna Biosciences
08-174
SYK [0.5]ULight-TK (24.6)
ULight-JAK-1 (20.0)
ULight-poly GAT (4.1)
Carna Biosciences
08-176
TEC [0.5]ULight-poly GT (25.4)
ULight-TK (18.6)
ULight-poly GAT (15.4)
Carna Biosciences
08-182
TIE2 [0.5]ULight-poly GT (19.3)
ULight-TK (17.7)
ULight-poly GAT (9.4)
Carna Biosciences
08-185
TNK1 [1]ULight-poly GAT (15.2)
ULight-poly GT (9.3)
ULight-TK (6.8)
Carna Biosciences
08-104
TRKA (NTRK1) [0.5]ULight-TK (23.2)
ULight-poly GAT (17.5)
ULight-poly GT (16.3)
Carna Biosciences
08-186
TRKB (NTRK2) [0.5]ULight-TK (28.0)
ULight-poly GAT (26.3)
ULight-poly GT (22.9)
Carna Biosciences
08-187
TRKC (NTRK3) [0.5]ULight-poly GT (34.1)
ULight-poly GAT (22.3)
ULight-TK (6.4)
Carna Biosciences
08-188
TXK [3]ULight-poly GAT (18.9)
ULight-poly GT (11.0)
ULight-TK (5.9)
Carna Biosciences
08-183
TYK2 [4]ULight-JAK-1 (15.2)
ULight-IRS-1 (7.3)
ULight-TK (7.3)
Carna Biosciences
08-147
TYRO3 [0.25]ULight-poly GT (21.5)
ULight-poly GAT (9.1)
ULight-TK (6.1)
Carna Biosciences
08-109
YES [0.25]ULight-TK (28.0)
ULight-poly GT (15.3)
ULight-poly GAT (14.0)
Carna Biosciences
08-175
ZAP70 [4]ULight-poly GAT (15.3)
ULight-poly GT (11.1)
ULight-TK (7.6)
Carna Biosciences
08-077
15
www.perkinelmer.com/lance
Kinase assay components:
Kinases: see table for concentration
ATP: 200 µM in kinase reaction
ULight-poly GT, ULight-poly GAT and ULight-IRS-1:100 nM in kinase reaction
ULight-JAK-1, ULight-TK and ULight-CDK1: 50 nM in kinase reaction
Europium anti-phospho-substrate antibodies: 2 nM in final volume
Kinase buffer:
50 mM HEPES pH 7.5
10 mM MgCl2
1 mM EGTA
2 mM DTT and 0.01% Tween 20
Kinase reactions were performed in 10 µL in white OptiPlates™-384 (cat# 6007290). After 2 h incubation at 23 °C, kinase reactions were stopped with EDTA (10 mM final concentration in 20 µL). Eu-anti-phospho antibodies were
added and incubated for one hour prior to detection with the EnVision®. All kinases were sourced from commercial vendors, and from commercially available lots.
LANCE Ultra products used for this study:
Catalog number
Phospho-Motif
ULight-poly GT
TRF0100
[EY(4:1)]n
ULight-poly GAT
TRF0101
[EAY(1:1:1)]n
ULight-IRS-1 (Tyr983) Peptide
TRF0120
SRGDYMTMQ
ULight-JAK-1 (Tyr1023) Peptide
TRF0121
DKEYYTVKD
ULight-CDK1 (Tyr15) Peptide
TRF0122
GEGTYVVY
ULight-TK Peptide
TRF0127
Y
Europium-anti-phosphotyrosine (PT66)
AD0068, AD0069
Y
KinaSelect TK kit*
TRF0301-D
Y
*contains ULight-TK Peptide, 0.5 nmole, Europium-anti-phosphotyrosine (PT66), 10 µg and Lance Detection Buffer 10X, 1.8 mL
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