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Home / Assay Kits

MicroMolar NADPH Assay Kit

SKU : BHE16300254

This product includes 1 ml of 10x Assay buffer, 0.1 ml of 20 x Reagent A, 0.1 ml of 20 x Reagent B, 0.5 ml of 10x Reagent C and 0.020 ml of 10 mM NADPH.

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Mfr.No.	NADPH100
Description	The assay measures micromolar concentrations of NADPH. NADPH is the reduced form of nicotinamide adenine dinucleotide phosphate (NADP). NADPH is an enzymatic cofactor for many reductases and acts as an electron donor. It is a product of many oxidases using NADP as an electron acceptor. Both NADPH and NADP are involved in many biosynthesis and degradation pathways. The MicroMolar NADPH Assay Kit (NADPH100) provides a convenient tool for sensitive detection of NADPH. The assay is based on detection of light absorbance at 590 nm and the assay process is completed within 20 min. It is in a 96-well plate format and can be used for high throughput screening of enzymes involving NADPH. The assay is compatible with most common buffers in biochemistry labs. It is not compatible with NADH or other reducing reagents such DTT. The MicroMolar NADPH Assay Kit (Catalog No. NADPH100) includes 1 ml of 10x Assay buffer, 0.1 ml of 20 x Reagent A, 0.1 ml of 20 x Reagent B, 0.5 ml of 10x Reagent C and 0.020 ml of 10 mM NADPH. It is for 100 assays in a 96-well plate format.
Concentration measurement	Biomolecules

Product Manual	Datasheet
Protocols Description	ASSAY PROTOCOL The following assay protocol is based on using a 96-well plate for the measurement. The sample volume is 100 µl and the final assay volume is 190 µl. It can be reformatted in a 384-well plate assays or using a cuvette by adjusting the volumes accordingly. STANDARD CURVE 1. Reagent dilution and sample preparation: Dilute the 20 x stock solutions 20-fold with water. Dilute the 10 x stock solutions 10-fold with water. Prepare 100 µl of NADPH solutions in the wells of a clear 96-well plate with a two-fold serial dilution from 100 µM to zero in 1 x assay buffer. 2. Detection: Into the 100 µl of the NADPH solutions, add 20 µl of 1 x Reagent A, then 20 µl of 1 x Reagent B. Incubate the mixture at room temperature for 10 min then add 50 µl of 1 x Reagent C and read the light absorbance at 590 nm. 3. Data Analysis: Plot the light absorbance Ac and the NADPH concentration [NADPH] to generate the linear standard curve. Ac = a [NADPH] + b Where the Ac values are from experimental data, the a and b values are from the linear fitting between the Ac values and the NADPH concentrations. UNKNOWN SAMPLES Follow the same procedure to measure the light absorbance Ac values from the unknown samples. Calculate the NADPH concentrations in the unknown samples using the Ac values from the unknown samples and the a and b values from the standard curve. [NADPH] = (Ac – b) / a

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SKU	Supplier No.	Size	Your price	Quantity
BHE16300254	NADPH100	100 assays	$302.17		Add to Cart

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