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Home / Assay Kits

E. coli BirA - for 100 assays

BHE16300166

This product includes 35 ul of 5 uM E. coli BirA.

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Product Manual Datasheet
Protocols Description Assay Protocol using E. coli Biotin Protein Ligase Assay Kit
The following assay protocol is based on the 384-well plate assay format. The reaction volume is 30 µl and the final assay volume is 75 µl. For 96-well plate assays, the reaction volume is 60 µl and the final assay volume is 150 µl. For detection using a cuvette, the reaction volume is 400 µl and the final assay volume is 1000 µl.

1. Reagent preparation:
For each 10 assay reactions,
(1) Prepare 297 µl of premix composed of 261 µl of H2O, 33 µl of 10 x Buffer, 3.3 µl of 100 x E. coli BirA and 3.3 µl of 100 x PPase.
(2) Prepare 33 µl of 10 x Enzyme substrate by mixing 3.3ml of 100 x Biotin, 3.3 µl of 100 x hydroxylamine, 3.5 µl of 100 x ATP with 23.1ml of water.
 
2. Reaction:
Mix 27 µl of the premix with 3 µl of the 10 x Enzyme substrate in each well. Incubate the reaction mixture at a temperature between 25 °C and 37 °C for 60 min.
 
3. Detection:
Add 45 µl of the Dye MPA3000 into the 30 µl of the reaction mixture. Incubate for 5 min. Measure the light absorbance at 650 nm.
SKU Supplier No. Size Your price Quantity
BHE16300166 BIRA-100KEC 100 assays $421.56
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