TECHNICAL DATA SHEET AlphaScreen® Research Reagents Caution: For Laboratory Use. A research reagent for research purposes only GST (Glutathione S-Transferase) Detection Kit Product No.: 6760603C/M/R Lot No.: 2025361 Material Provided Format: 6760603C 6760603M 6760603R 500 Assay Points 10000 Assay Points 50000 Assay Points Note: The number of assay points is based on a final bead concentration of 20 µg/mL in a 25 µL/well reaction volume. Manufacturing Date: May 6, 2015 Kit Components: Component Anti-GST (Glutathione S-Transferase) Acceptor Beads at 5 mg/mL in 25 mM Hepes, 100 mM NaCl, 0.05% Proclin-300, pH 7.4 Streptavidin Donor Beads at 5 mg/mL in 25 mM Hepes, 100 mM NaCl, 0.05% Proclin-300, pH 7.4 Biotinylated-GST at 0.5 µM in 25mM Hepes, 100mM NaCl, 0.05% Proclin-300, 0.05% Tween 20, pH 7.4 10x Buffer : 250 mM Hepes, 1 M NaCl, 0.05% Proclin-300, pH 7.4 6760603C 6760603M 6760603R 1 x 50 µL 1 x 1 mL 1 x 5 mL (6760106) (6760107) (6760107B) 1 x 50 µL 1 x 1 mL 1 x 5 mL (6760001) (6760002) (6760002B) 1 x 50 µL 1 x 50 µL 1 x 50 µL (6760304) (6760304) (6760304) 1 x 1.5 mL 1 x 1.5 mL 1 x 1.5 mL (6760020G) (6760020G) (6760020G) Product Information Antibody/Protein: The GST antibody is a goat polyclonal antibody that recognized Glutathione STransferase [Schistosoma japonicum]. Stability: This product is stable for at least 12 months from the manufacturing date if used and stored under recommended conditions. Storage Conditions: Store undiluted at 4°C protected from light. Freeze-thaw is not recommended and can cause the beads to form aggregates. Recommended use: AlphaScreen® Donor beads are light-sensitive. All Alpha assays using the Donor beads should be performed under subdued laboratory lighting (< 100 lux). Green filters (LEE 090 filters (preferred) or Roscolux filters #389 from Rosco) can be applied to light fixtures. TDS-6760603C/M/R-02 Page 1 of 3 ® For additional information on running AlphaScreen assays or on potential interfering compounds, please visit our website: www.perkinelmer.com/AlphaTech Quality Control ® AlphaScreen maximum signal, minimum signal and EC50 are determined using a biotinylated-GST titration assay ® performed on an EnVision HTS Alpha detection instrument. We certify that these results meet our requirements. TEST RESULTS Maximum signal Minimum signal EC50 303179 1172 0.40 nM Recommended Assay Conditions Note: This protocol provides a method to verify kit performance and is not representative of an assay. Sufficient biotinylated-probe and 10x buffer is provided to perform 3 titration curves in triplicate as described. ® 1x Buffer: Add 500 μL 10x buffer to 4.5 mL Milli-Q H2O (or equivalent). Add 5 mg BSA (0.1% final concentration) and adjust pH to 7.4. Acceptor Beads: Add 5 μL Anti-GST (Glutathione S-Transferase) Acceptor beads to 245 μL 1x buffer. Donor beads: Add 5 μL Streptavidin Donor beads to 245 μL 1x buffer. Biotinylated-probe: From the 0.5 µM biotin-GST, prepare a ½ log dilution series (50 nM to 5 pM) in 1x buffer. Include a buffer only control. Titration Protocol: To a white opaque 384-well Optiplate: 1- Add 5 μL biotinylated-GST dilutions (from lowest to highest concentration). 2- Add 10 μL of 1x buffer 3- Add 5 μL of Anti-GST (Glutathione S-Transferase) Acceptor beads. Incubate in the dark at room temperature for 30 minutes. 4- Add 5 μL of Streptavidin Donor beads. ® Incubate in the dark at room temperature for 60 minutes and analyze on your AlphaScreen detection reader. TDS-6760603C/M/R-02 Page 2 of 3 AlphaScreen signal (counts) Product Typical Data 500000 400000 300000 200000 100000 0 - -13 -12 -11 -10 -9 -8 log [b-GST] (M) Figure 1: Biotinylated-probe titration assay 384-well biotinylated-GST titration curve (25 μL final volume; Reader: Envision HTS Alpha). ® Note: AlphaScreen signal will vary depending on instrument detection protocol, incubation temperature and incubation time. Suggested Materials and Instrumentation Please visit our website www.perkinelmer.com/AlphaTech This product is not for resale or distribution except by authorized distributors. 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