Carbonic Anhydrase 2 Human Enzymatic LeadHunter Assay - TW

Procedure Summary:

Human erythrocyte carbonic anhydrase II is used. Test compound and/or vehicle is preincubated with 15 μg/ml enzymeǂ in 50 mM MOPS solution pH 7.5 for 15 minute at 25⁰C. The reaction is initiated by addition of 500 µM 4-nitrophenyl acetate and is incubated for 30 minutes. The plate is read on the spectrophotometer (Absorbance: 400 nm). Compounds are screened at 10 μM.

Substrate:

500 µM 4-Nitrophenyl acetate (4-NPA)

Incubation:

30 min at 25°C

Control Inhibitor:

Acetazolamide

Criteria For Significance:

≥ 50% of max stimulation or inhibition

Test Concentration / Dose:

Suggested concentration for initial testing is 10 uM

Test Sample Requirements:

Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).

Minimum Order Quantity:

2