S. pneumoniae MurF Assay Kit Plus-100

Mfr.No.	MURF100KN
Description	MurF is the enzyme that catalyzes the last step in synthesis of UDP-MurNAc-pentapeptide in the pathway of peptidoglycan biosynthesis in bacteria. It is an essential enzyme and attractive target for anti-bacterial drug discovery. MurF adds a dipeptide D-Ala-D-Ala onto the MurE product UDP-MurNAc-tripeptide. The ligation reaction is coupled to the hydrolysis of ATP forming ADP and inorganic phosphate. The S. pneumoniae MurF Assay is based on measurement of the inorganic phosphate generated from the MurF reaction. The inorganic phosphate is detected by light absorbance at 650 nm. The assay reactions and detection can be performed by using 384-well or 96-well assay plates. Alternatively, the assay reaction can be carried out in Eppendorf tubes and the signal is measured using a cuvette. The high throughput assay can be used for screening inhibitors of E.coli MurF in drug discovery research. It may also be used for characterization of E.coli MurF. The S. pneumoniae MurF Assay Kit Plus-100 (Catalog No. MURF100KN) contains the reagents for 100 assays in a 384-well plate assay format including 400µl of 10 x Buffer, 33µl of 100 x UDP-MurNAc-tripeptide (UMAG-DAP), 33µl of 100 x D-Ala-D-Ala, 33µl of 100 x ATP , 33µl of 100 x S. pneumoniae MurF (2000 nM) and 5 ml of Dye MPA3000 for phosphate detection.
Drug target assays	Antibacterial drug targets

Product Manual	Datasheet
Protocols Description	ASSAY PROTOCOL The following assay protocol is based on the 384-well plate assay format. The reaction volume is 30µl and the final assay volume is 75µl. For 96-well plate assays, the reaction volume is 60µl and the final assay volume is 150µl. For detection using a cuvette, the reaction volume is 400µl and the final assay volume is 1000µl. 1.Reagent preparation: For each 10 assay reactions, (1) Prepare 297µl of premix composed of 261µl of H2O, 33µl of 10 x Buffer and 3.3µl of 100 x S. pneumoniae MurF. (2) Prepare 33µl of 10 x Enzyme substrate by mixing 3.3µl of 100 x UMAG-DAP, 3.3µl of 100 x D-Ala[1]D-Ala, 3.3µl of 100 x ATP, and 23.1µl of water. 2. Reaction: Mix 27µl of the premix with 3µl of the 10 x Enzyme substrate in each well. Incubate the reaction mixture at 37°C for 60 min. 3. Detection: Add 45µl of the Dye MPA3000 into the 30µl of the reaction mixture. Incubate for 5 min. Measure the light absorbance at 650 nm.

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SKU	Supplier No.	Size	Your price	Quantity
BHE16300062	MURF100KN	100 assays	$613.14		Add to Cart