MMP-3 [3H] enzyme assay
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1536-well assay protocol>
Reagent usage in 384- and 1536-well formats>
Typical data>
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Assay concept
[3H]Y-A-R-G-A-I-G-L-P-A-AHA-biotin Streptavidin-coated beads
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1536-well assay protocol
- 10 µl 50 mM Tris pH 7.5, 10 mM CaCl2, 0.1% (w/v) BSA
- 10 µl labelled peptide (35 000 cpm)
- 10 µl MMP-3 (concentration unknown) or 10 µl water for no enzyme controls
- Incubate for 40 minutes at 37oC
- 20 µl streptavidin polystyrene (RPNQ0261) or streptavidin yttrium oxide (RPNQ0271) bead (0.25 µg) in stop reagent containing EDTA (0.56 M, pH 8)
- Incubate for 30 minutes at room temperature
- Image for 10 minutes
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Reagents usage in 384- and 1536-well formats
Assay components | 384-well | 1536-well |
Assay volume | 50 µl | 3 µl |
Bead | 250 µg | 25 µg |
Enzyme | 10 µl | 1 µl |
Substrate | 35 000 cpm | 4 000 cpm |
Substrate amount | 10 µl | 1 µl |
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Typical data
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| Substrate titration in 384-well microplate | Enzyme titration in 384-well microplate |
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Ordering information
The following products were used in this assay and can be purchased online
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