• Product name
    NADP/NADPH Assay Kit
    See all NADP/NADPH kits
  • Detection method
  • Sample type
    Tissue Extracts, Cell Lysate
  • Assay type
  • Assay time
    2h 00m
  • Product overview

    NADP/NADPH Assay Kit (ab65349) provides a convenient tool for sensitive detection of the intracellular nucleotides: NADP, NADPH and their ratio. Assays of nicotinamide nucleotides are of continual interest in the studies of energy transforming and redox state of cells or tissue.

    The enzymes in the system specifically recognize NADP/NADPH in an enzyme cycling reaction (It does not recognize NAD+/NADH). There is no need to purify NADP/NADPH from sample mix. The enzyme cycling reaction significantly increases detection sensitivity. Results can be quantified using plate reader at OD450nm.
    Visit our FAQs page for tips and troubleshooting.


  • Notes

    If you would like to use a fluorometric reading, please refer to NADP/NADPH Assay Kit (Fluorometric) (ab176724).

  • Tested applications
    Suitable for: Functional Studiesmore details
  • Platform


  • Storage instructions
    Store at -80°C. Please refer to protocols.
  • Components Identifier 100 tests
    NADP Cycling Buffer WM 1 x 15ml
    NADP Cycling Enzyme Mix Green 1 x 0.2ml
    NADP/NADPH Extraction Buffer NM 1 x 50ml
    NADPH Developer Purple 1 vial
    NADPH Standard (MW:833.36) Yellow 1 x 166.7µg
    Stop Solution Red 1 x 1.2ml
  • Research areas
  • Relevance
    NADP (Nicotinamide adenine dinucleotide phosphate) is a coenzyme composed of ribosylnicotinamide 5-phosphate (NMN) coupled by pyrophosphate linkage to the 5-phosphate adenosine 2,5-biphosphate. It serves as an electron carrier in a number of reactions, being alternately oxidised (NADP+) and reduced (NADPH). The oxidative phase of the pentose phosphate pathway is the major source of NADPH in cells, producing approxiamtely 60% of the NADPH required. NADPH provides the reducing equivalents for biosynthetic reactions and the oxidation-reduction involved in protecting against the toxicity of ROS, allowing the regeneration of GSH. NADPH is also used for anabolic pathways, such as lipid synthesis, cholesterol synthesis and fatty acid chain elongation.


Our Abpromise guarantee covers the use of ab65349 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Functional Studies Use at an assay dependent dilution.


  • NAPDH/NADP+ ratio was determined using ab65349 in stable HSPB1- knockdown U87 cells. 

  • Standard curve with background signal subtracted (duplicates; +/- SD).

  • Total NADP and NADPH (tNADP) or NADPH alone measured in RAW cell lysates (duplicates +/- SD).

  • Measurement of NADPt and NADPH in rat liver lysate (20 μg). Assays were performed using the kit protocol.

  • Measurement of NADPt and NADPH in HeLa cell lysate (80 μg). Assays were performed using the kit protocol.

  • Typical standard curve for ab65349.



This product has been referenced in:
  • Zhu H  et al. IDH1 R132H Mutation Enhances Cell Migration by Activating AKT-mTOR Signaling Pathway, but Sensitizes Cells to 5-FU Treatment as NADPH and GSH Are Reduced. PLoS One 12:e0169038 (2017). Functional Studies . Read more (PubMed: 28052098) »
  • Dey P  et al. Genomic deletion of malic enzyme 2 confers collateral lethality in pancreatic cancer. Nature 542:119-123 (2017). Functional Studies . Read more (PubMed: 28099419) »

See all 22 Publications for this product

Customer reviews and Q&As

We highly recommend to follow storage instructions on the individual datasheets.  For ab65349 kit it should be stored at -80oC when it is delivered and for short term storage.

It is also worth noting that the components of some kits, once reco...

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I am sorry we have not yet tested ab65349 NADP/NADPH Assay Kit in serum samples so we do not have laboratory information to provide to help answer your question. All tested sample types covered by the Abpromise guarantee are listed on the individu...

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Thank you for your enquiry.
I have now heard back from the laboratories. They have confirmed that they have not tested this kit with samples stored in RNAlater but you can remove the RNA later solution and use the extraction buffer to prepare the s...

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1. Some liquid loss during heating (can happen even in tubes if the liquid is not spun down). Be sure to cool and spin down the tubes following decomposition.

2. Endogenous enzymes converting to NADPH if the samples are not deproteinize...

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NADPH oxidase is typically a complex made up of a cluster of proteins that donate an electron from NADPH to molecular oxygen (O2) to produce superoxide (O2-). This enzyme activity is not measured by this kit directly.

Due to NADPH ox...

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Below is our recommended procedure. The key is to keep your samples on ice during homogenization to avoid heating and degradation of the analyte and to ensure lysis under the microsope before proceding with the assay.

General Sample prepa...

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If you suspect insufficient lysis, I recommend taking a small volume and visualizing under a microscope to ensure that the cell membrane has been lysed. If not, you can try an additional freeze-thaw cycle, or try homogenizing with a dounce homogenizer....

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One modification to the protocol that may help is the following:
Seal the plate and then heat at 60C for 30 mins. During this time evaporation reduces the volume and hence makes the solution more concentrated which is why often the NADPH OD is gre...

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I double checked and unfortunately, have confirm that we do not have a kit for measuring NADPH oxidase in our catalog at the moment. We do offer ab65349 which is a NADP/NADPH Assay Kit:


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We recommend using the buffers provided with the specific kits for our assays. So, unfortunately we don’t suggest using the same buffer for both assays (ab65348 and ab65349).

1-10 of 61 Abreviews or Q&A


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