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NADH/NAD Quantification Kit (Colorimetric) (ab65348) provides a convenient tool for sensitive detection of the intracellular nucleotides: NADH, NAD and their ratio. Assay of nicotinamide nucleotides is of continual interest in the studies of energy transforming and redox state of cells or tissues.
The NAD Cycling Enzyme Mix in the kit specifically recognizes NADH/NAD in an enzyme cycling reaction. There is no requirement to purify NADH/NAD from samples. The reaction specifically detects NAD and NADH, but not NADP nor NADPH. The enzyme cycling reaction significantly increases the detection sensitivity and specificity. NADt (NAD and NADH) or NADH can be easily quantified by comparing with standard NADH.
Visit our FAQs page for tips and troubleshooting.
Review our Metabolism Assay Guide to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader.
If you would like to use a fluorometric reading, please refer to NAD/NADH Assay Kit (Fluorometric) (ab176723).
|NAD Cycling Buffer||NM||1 x 15ml|
|NAD Cycling Enzyme Mix||Green||1 vial|
|NADH Developer||Purple||1 vial|
|NADH Standard (MW:663.4)||Yellow||1 x 200nmole|
|NADH/NAD Extraction Buffer||NM||1 x 50ml|
|Stop Solution||Red||1 x 1.2ml|
Our Abpromise guarantee covers the use of ab65348 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Functional Studies||Use at an assay dependent dilution.|
Image obtained from Ren JG et al; PLOS one, 2010; 5(9): e12520 (DOI:10.1371/journal.pone.0012520)
NAD and NADH (tNAD) or NADH alone measured cell lysates. 5e6 cells were lysed in 1 mL, spin filtered, and tested neat or 1/5 (duplicates +/- SD).
Standard curve with background signal subtracted (duplicates; +/- SD).