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[33P] MAP kinase enzyme assay
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Assay concept>
1536-well assay protocol>
Reagent usage in 96-, 384- and 1536-well formats>
Typical data>
Ordering information>
Assay concept
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1536-well assay protocol
1. | 4 µl reaction mix:
0.5 µl reaction buffer containing 12.5 pmoles MBP substrate,
1 µl [33P]ATP (AH9968)
2.5 µl water
1 µl Erk1 MAPK enzyme |
2. | Incubate at 370C in a humidified incubator for 30 minutes |
3. | 3 µl stop mix:
1.33 µl stop solution plus ATP
0.5 µl polystyrene streptavidin (RPNQ0261) or yttrium oxide
(RPNQ0271) bead
1.17 µl PBS |
4. | Image for 5 minutes |
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Reagent usage in 96-, 384- and 1536-well formats
Assay components | 96-well | 384-well | 1536-well |
Assay volume | 250 µl | 75 µl | 7 µl |
Bead | 0.5 µg | 0.25 µg | 0.05 µg |
Enzyme amount | 1 µg | 0.5 µg | 0.1 µg |
Substrate | 2.5 mM | 1.66 mM | 3.4 mM |
Substrate amount | 125 pmoles | 50 pmoles | 10 pmoles |
ATP | 1 mM | 0.83 mM | 1.7 mM |
ATP amount | 50 pmoles | 25 pmoles | 5 pmoles |
[33P]ATP | 0.5 µCi | 0.1 µCi | 0.02 µCi |
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Typical data
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Ordering information
The following products were used in this assay and can be purchased online
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