Concentration assays for NADPH / Coenzyme A (CoA) / Glutathione (GSH)

 

 

 

1.  NADPH

 

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 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.

 

 

 

 

 

 

 

 

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, 1 ml of 10x Reagent C and 0.020 ml of 10mM NADPH. It is for 100 assays in a 96-well plate format.

 

 

 

 

 

 

 

2.  Coenzyme A (CoA)

 

The Coenzyme A Assay Kit is for measurement of micromolar concentrations of Coenzyme A (CoA). 

The assay is based on the principle that the thiol group of CoA interacts with the CAK reagent and enhances the fluorescence intensity at 535 nm (excitation at 485 nm). Acetyl-CoA does not interact with the CAK reagent.

 

 

 

 

 

 

 

Coenzyme A Assay Kit (Catalog No CAK1000)

The assay reagent is sufficient for measurement of 1000 samples using 384-well plates. The assay kit can be used for measurement of other thiols.

The assay is compatible with HEPES buffer, low concentrations of non-ionic detergent (<0.01%), sodium and potassium salt, MgCl2 (< 5 mM), CaCl2 (<5 mM) and phosphate (< 1 mM). The assay is not compatible with ammonium, Tris buffer or other primary amine buffers. It is not compatible with amino acids, DTT or EDTA.

 

 

 

 

 

3.  Glutathione (GSH)

 

Glutathione is an essential antioxidant in the cells that prevents damage of biomolecules due to oxidation. Glutathione is also a popular ingredient in pharmaceutical products. The MicroMolar Glutathione Assay Kit is for measurement of micromolar concentrations of glutathione. 

The assay is based on increase of the fluorescence intensity (emission 535 nm, excitation 485 nm) of the kit fluorescence dye MAA upon binding to glutathione. The assay detects reduced glutathione (GSH) at a much higher sensitivity than the oxidized form (GSSG).

 

 

 

 

 

 

 

MicroMolar Glutathione Assay Kit (Catalog No MGA200)

 

 

Includes 500 μl of 10 x MAA dye. It is for 200 assays using 96-well plates. The assay kit can be used for measurements glutathione concentrations in pharmaceutical products, biochemical reactions or other samples. The assay is compatible with HEPES buffer, low concentrations of non-ionic detergent (<0.01%), MgCl2 (< 5 mM), CaCl2(<5 mM), EDTA (< 1mM) and phosphate (< 1 mM). It is not compatible with thiol compounds such as DTT and cysteine. It is not compatible with samples with histidine.