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E. coli D-Alanine: D-Alanine Ligase Assay Kit Plus-100

BHE16300043

This product includes the reagents for100 assays in a 384-well plate assay format including 0.400 ml of 10 x Buffer, 0.035 ml of 100 x Enzyme Substrate, 0.035 ml of 100 x E. coli D-Ala:D-Ala ligase (200 nM) and 5 ml of Dye for phosphate detection.

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Product Manual Datasheet
Protocols Description ASSAY PROTOCOL
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 and 3.3 µl of 100 x E. coli D[1]Ala:D-Ala ligase.
(2) Prepare 33 µl of 10 x Enzyme substrate by mixing 3.3µl of 100 x Enzyme substrate with 29.7µl 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 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
BHE16300043 DDA100KE 100 assays $613.14
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