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Ultra-sensitive High-throughput Protease Assay Kit

BHE16300165

This product includes the 5 ml of 10 x assay buffer and 50 ul of 1000 x substrate for 1000 assays of proteases in a 384-well plate format.

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+1 866.986.9598
Email
[email protected]
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Product Manual Datasheet
Protocols Description Assay Protocol
1. Reagent preparation
Thaw the 1000 x protease substrate from the frozen stock. Mix the stock solution well by shaking or vortex. Dilute the 1000 x protease substrate 100-fold with water to make the 10 x protease substrate.

2. Assay
In a standard black 384-well plate (Corning plate 3571, for example), mix 5 µl of 10 x buffer, 40 µl of the protein sample and 5 µl of 10 x protease substrate. Incubate the reaction mixture at 37°C for 2 hours. Measure the fluorescence intensity at 535 nm (excitation at 485 nm).
 
Note: It is a continuous assay. The fluorescence signal can be measured at different time points. A longer incubation including overnight or days allows to detect a lower concentration of protease.
 
Assay Protocol for enzyme inhibition
Enzyme inhibition IC50 can be measured using the 384-well or 96-well plate assay format. Typically 50 x stock solutions with a 2-fold serial dilution in water or DMSO are prepared. For 96-well plate assays, 4 µl of the 50 x inhibitor is mixed with 176 µl of the assay reaction mixture composed of the buffer and enzyme for 5 min. Then 20 µl of 10 x substrate is added. At the end of the reaction, the fluorescence intensity is measured.
SKU Supplier No. Size Your price Quantity
BHE16300165 UPA1000 1000 assays $289.02
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