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Home / Assay Kits

Bacterial RNA Polymerase Pyrophosphate Assay Kits

SKU : BHE16300148

This product includes 400 ul of Buffer RP, 33 ul of 100 x DNA, 33 ul of 100 x NTP mix, 5 ul of 1000 x pyrophosphatase and 4500 ul of dye for 100 assays of E. coli RNA polymerase reactions in a 384-well assay format. The assay This product includes all reagents except the enzyme.

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Mfr.No.	RPAP100K, RPAP100KE, PPA1000, RPAP100KSE
Description	The bacterial RNA polymerase is responsible for biosynthesis of mRNA, tRNA and rRMA in the cells. The holoenzyme of bacterial RNA polymerases contains multiple subunits with a molecular mass of about 400 kDa. The bacterial RNA Polymerase Pyrophosphate Assay Kit is based on measurement of the pyrophosphate generated in the RNA polymerase reaction using a double strand DNA template. The pyrophosphate is converted to phosphate by pyrophosphatase and the phosphate is detected by the phosphate detection reagent (catalog number MPA3000). The assay can be performed in a 384-well plate format for high throughput screening of RNA polymerase inhibitors. For assay kits measuring RNA formation, please see Related Products below. The Bacterial RNA Polymerase Pyrophosphate Assay Kit (Catalog number RPAP100K) includes 400 µl of Buffer RP, 33 µl of 100 x DNA, 33 µl of 100 x NTP mix, 10 µl of 1000 x pyrophosphatase and 4500 µl of dye for 100 assays of E. coli RNA polymerase reactions in a 384-well assay format. The assay kit includes all reagents except the enzyme.
Drug target assays	Antibacterial drug targets

Product Manual	Datasheet
Protocols Description	Assay Protocol 1. Reagent preparation: 1x assay buffer: dilute the 10 x Buffer RP 10-fold with water. 10 x DNA: dilute the 100 x DNA 10-fold with water. 10 x enzyme: dilute the 100 x RNA polymerase (Catalog #: RNAP-100EC) 10-fold with the 1 x assay buffer. 10 x NTP mix: dilute the 100 x NTP mix 10-fold with water. 10 x pyrophosphatase: dilute the 1000 x pyrophosphatase (100 U/ml) 100-fold with the 1 x assay buffer. 2. Reaction: A transparent 384-well plate (Matrix 4310, for example) is used for the assay. The total volume of each reaction mixture is 30 µl including 15 µl of H2O, 3 µl of 10 x Buffer RP, 3 µl of 10 x DNA template, 3 µl of 10 x enzyme, 3 µl of 10 x pyrophosphatase, 3 µl of 10 x NTP mix. Incubate the reaction mixture at 37°C for 60 min. 3. Detection: Add 40 µl of the dye (MPA3000) into the 30 µl of the reaction mixture. Incubate for 3 min. Measure the light absorbance at 650 nm.

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Combination SKU	Supplier No.	Specifications	Your price	Quantity
BHE16300148-1	RPAP100K	Bacterial	$365.40		Add to Cart
BHE16300148-2	RPAP100KE	E. coli	$613.14		Add to Cart
BHE16300148-3	PPA1000	Pyrophosphate Assay Reagent	$489.27		Add to Cart
BHE16300148-4	RPAP100KSE	S. aureus	$613.14		Add to Cart

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