| Measurement of Activity: |
PARG activity is measured by the loss of biotinylated PAR from histones attached to the wells of a 96 well plate. |
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| Assay Conditions: |
The reaction (50 µl) takes place in wells of a 96 well plate to which is attached biotin-conjugated-poly(ADP) ribosylated histones, and consists of 20 mM Potassium Phosphate (pH 7.4), 50 mM KCl, 0.1 mg/ml BSA, 0.1% Triton X-100, and serial dilutions of PARG. Incubation is for 30 minutes at room temperature. The loss of biotinylated poly(ADP-ribose) is quantified by incubation with Streptavidin-HRP followed by addition of a colorimetric HRP substrate and measurement of the absorbance at 450 nm. A PARG concentration of 10.25 ng/ml will cleave 50% of the PAR under these conditions. |
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| Storage: |
Stable for at least one year when stored at -20°C. |
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| Storage Buffer: |
20 mM KPO4 (pH 7.2), 50 mM KCl, 0.1 mg/ml BSA, 0.1% Triton® X-100, 1mM DTT, 50% Glycerol. |
