| Solution Information | help | |
| Enzyme: | Serine/threonine-protein kinase PLK1 | |
| inhibitor: | BDBM41778 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Protocol for Redox Cycling H2O2 Generation Assay A simple colorimetric 384-well format H2O2 generation assay has been developed and adapted by the PMLSC from a tube based assay to measure H2O2 production by chemically elicited peritoneal macrophages that had been plated in either 3-cm-diameter plastic tissue culture petri dishes, or in 24-well microtiter plates. The assay is based on the ability of horse radish peroxidase (HRP) to catalyze the oxidation of phenol red by H2O2, producing a change in it's absorbance at 610 nm after the assay reaction has been terminated and adjusted to alkaline pH by the addition of NaOH (Pick & Keisari (1981), Cell. Immunol. 59: p301). In the current format, the assay involves three liquid transfer steps into a 384-well plate, 20 uL each of compounds/controls, DTT, and phenol red-HRP to give a final assay volume of 60 uL. After a 30 minute incubation period at ambient temperature the assay is terminated by the addition of 10 uL of 1N NaOH an | |
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