| Assay Method Information | |
| | Assay Protocol for Determination of IC50 Against Human CD38 |
| Description: | The CD38 Inhibitor Screening Assay Kit (BPS Bioscience, Catalog #: 79287) is designed to measure the glycohydrolase activity of CD38 for screening and profiling applications.CD38 hydrolase buffer (4×) was thawed on ice and 1×CD38 hydrolase buffer was prepared by diluting 4×CD38 hydrolase buffer with water. CD38 hydrolase enzyme was diluted to 0.5 ng/ml with 1×CD38 hydrolase buffer (10 ng/well). Master mix was prepared by mixing equal volumes of water and CD38 hydrolase buffer (4×) and 10 μl of the mix was added to all the wells. Following master mix addition, 10 μl of the inhibitors and 20 μl of CD38 hydrolase enzyme were added to the wells. For wells designated blank, 10 μl and 20 μl of 1× CD38 hydrolase buffer was added respectively in place of CD38 enzyme and inhibitors. The plate was covered and incubated 30 min at room temperature with slow shaking at 400 rpm. After 30 min incubation, the plate was removed and 10 μl of diluted F-NAD substrate was added to all the wells. The plate is incubated for 10 min, spun at 1200 rpm for 30 secs to remove bubbles and read in Tecan Spark using a fluorometer setting of excitation 300 nm and emission 410 nm. The reading for the blank was subtracted from all the values. |
| Affinity data for this assay | |
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