| Assay Method Information | |
| | PTP activity assay |
| Description: | The enzyme activity was measured using p-nitrophenyl phosphate (pNPP) as substrate in a 96-well plate and by detecting the absorbance at 405 nm for the amount of produced p-nitrophenol. Purified human recombinant PTP1B or TCPTP (0.05 µg) in 50 µL buffer containing 50 mM citrate (pH 6.0), 0.1 M NaCl, 1 mM EDTA, and 1 mM DTT and test compounds were added to each well. Each mixture was pre-incubated for 15 min at room temperature, followed by the addition of 50 µL of reaction buffer containing 2 mM pNPP and incubated at 37°C for 30 min. |
| Affinity data for this assay | |
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