Assay Method Information

Assay Name:  Fluorescent-based In Vitro Assa
Description:  The assay was performed in Optiplate 96-wells black plates, with each reaction well containing a mixture of 25 mM sodium acetate buffer pH 4.5 and a fixed amount of protein (2 μg) in a volume of 85 μL. After 30 mins of pre-incubation with test compounds (diluted 20× from DMSO stock solutions at different concentrations), the fluorogenic probe was added (diluted 20× from EtOH stock solution, final concentration 5 μM). After incubation for 3 hours at 37° C., the reactions were stopped with 50 μL of methanol and 100 μL of a 2.5 mg/mL NalO4 fresh solution in 100 mM Glycine/NaOH pH 10.6. The plate was incubated at 37° C. for 2 hours in the dark and fluorescence intensities were measured at excitation/emission wavelengths of 360/446 nm. Negative control samples consisted of the same incubation mixture in the absence of protein extracts.
Affinity data for this assay
 

If you find an error in this entry please send us an E-mail