Antibody - PerkinElmer

LANCE® Ultra Research Reagents
Caution: For Laboratory Use. A product for research purposes only
Eu-W1024-labeled Anti-phospho-Histone H3 (Ser10)
Product No.: TRF0210-D / TRF0210-M
Lot No.: 1606201
Material Provided
10 µg (1 562 assay points*)
100 µg (15 625 assay points*)
*Assuming 40 fmol/ assay point
100 µL (TRF0210-D) or 1 mL (TFR0210-M)
Manufacturing Date:
March 12, 2012
Product Information
Europium-labeled mouse monoclonal antibody recognizing phospho-Ser10 in
human Histone H3.
Molecular Weight:
160 000
Storage Buffer:
50 mM Tris-HCl (pH 7.4), 0.9% NaCl, 0.1% BSA and 0.05% sodium azide as
This product is stable for at least 12 months from the manufacturing date when
stored in its original packaging and the recommended storage conditions.
Storage Conditions:
Store at 4°C
Safety Note:
The storage buffer contains sodium azide (NaN3) as a preservative. Disposal of all
waste should be in accordance with local regulations.
Quality Control
The QC release specifications are based on spectrophotometric analysis of the labeled antibody. We certify that
these results meet our quality release criteria.
Labeling Ratio:
Page 1 of 3
6.9 (Eu/Ab)
100 µg/mL (0.625 µM)
Recommended Assay Conditions
AURORA B kinase: ATP titration
Reagent Preparation:
Prepare 1X Kinase Assay Buffer: 50 mM HEPES pH 7.5, 1 mM EGTA, 10 mM MgCl2, 2 mM DTT and 0.01%
Prepare a 2X Aurora B solution: dilute enzyme to a concentration of 0.2 nM in 1X Kinase Assay Buffer. Keep
on ice.
Prepare a 4X ULight-Histone H3 (Thr3/Ser10) Peptide solution: dilute ULight-Histone H3 (Thr3/Ser10) Peptide
to a concentration of 200 nM in 1X Kinase Assay Buffer.
Prepare a 4X ATP solution: dilute ATP to concentrations ranging from 40 nM to 4 mM (serial half-log dilutions)
in 1X Kinase Assay Buffer. Keep on ice.
Prepare 1X Detection Buffer: dilute 0.5 mL of 10X Detection Buffer with 4.5 mL of H2O.
Prepare a 4X Stop solution*: dilute EDTA to a concentration of 40 mM in 1X Detection Buffer.
Prepare a 4X Detection Mix: dilute Europium-anti-phospho-Histone H3 (Ser10) Antibody to a concentration of
8 nM in 1X Detection Buffer.
Pipet 5 µL of 2X Aurora B solution into a 384-well white OptiPlate-384 (0.1 nM final concentration).
Add 2.5 µL of 4X ULight-Histone H3 (Thr3/Ser10) Peptide solution (50 nM final concentration).
Add 2.5 µL of 4X ATP solution (10 nM to 1 mM final concentrations).
Cover plate with TopSeal-A and incubate 60 min at 23°C.
Add 5 µL of 4X Stop solution* and incubate 5 min at 23°C.
Add 5 µL of 4X Detection Mix (2 nM Europium-anti-phospho-Histone H3 (Ser10) Antibody final concentration).
Cover plate with TopSeal-A and incubate 60 min at 23°C.
Remove TopSeal-A and read in TR-FRET mode at 665 nm (excitation at 320 or 340 nm).
*Alternatively, the Stop solution and Detection Mix can be premixed and added together to the kinase reaction.
Page 2 of 3
Typical ATP Titration Data
LANCE Signal (665 nm)
Aurora B kinase assay using ULight-Histone H3 (Thr3/Ser10) Peptide
and Eu-anti-phospho-Histone H3 (Ser10) Antibody
obtained using the EnVision® Multilabel Reader:
Km app = 1.8 M
Log [ATP] (M)
Suggested Materials
Cat. No.
Substrate: ULight™- Histone H3 (Thr3/Ser10) Peptide
Antibody: Eu- anti-phospho-Histone H3 (Ser10) Antibody
Kinase: Aurora B
Carna Biosciences
Detection Buffer: LANCE Detection Buffer, 10X
Plate: OptiPlate™-384, white
TopSeal™: TopSeal-A
Please visit our website for additional resource:
This product is not for resale or distribution except by authorized distributors.
LANCE® products may be covered by one or more the following patents: US 5,202,423 and US 5,324,825
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