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AB65349

NADP/NADPH Assay Kit

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(1 Review)

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(190 Publications)

NADP/NADPH Assay Kit ab65349 provides a convenient tool for sensitive detection of the intracellular nucleotides: NADP, NADPH and their ratio. Readout on any colorimetric (450 nm) plate reader.

- Cited in >140 publications
- Complete kit including standard curve for quantitation
- Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
7 Images
Functional Studies - NADP/NADPH Assay Kit (AB65349)
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Supplier Data

Functional Studies - NADP/NADPH Assay Kit (AB65349)

Typical standard curve for ab65349.

Functional Studies - NADP/NADPH Assay Kit (AB65349)
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Lab

Functional Studies - NADP/NADPH Assay Kit (AB65349)

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

Functional Studies - NADP/NADPH Assay Kit (AB65349)
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Lab

Functional Studies - NADP/NADPH Assay Kit (AB65349)

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

Functional Studies - NADP/NADPH Assay Kit (AB65349)
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Supplier Data

Functional Studies - NADP/NADPH Assay Kit (AB65349)

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

Functional Studies - NADP/NADPH Assay Kit (AB65349)
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Supplier Data

Functional Studies - NADP/NADPH Assay Kit (AB65349)

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

Functional Studies - NADP/NADPH Assay Kit (AB65349)
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PubMed

Functional Studies - NADP/NADPH Assay Kit (AB65349)

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

Image from Hongxing Y et al., PLoS One 11(10), Fig 2a. Doi:10.1371/journal.pone.0164285 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

Schematic Diagram - NADP/NADPH Assay Kit (AB65349)
  • Schematic Diagram

Supplier Data

Schematic Diagram - NADP/NADPH Assay Kit (AB65349)

Representative image of NADP/NADPH Assay Kit ab65349

Components shown from left to right :

- Developer Solution II

- NADPH Standard

- Stop Solution II

- NADP Cycling Enzyme Mix

- NADP Cycling Buffer

- Extraction Buffer II

Note : The vial labels shown in this image use generic names for illustrative purposes only and may not exactly match the specific component names included in the kit.

Note : Colors of solutions in image may not precisely match the shade of colors in the actual kit.

Key facts

Detection method

Colorimetric

Sample types

Tissue Extracts, Cell Lysate

Assay type

Quantitative

Assay time

2h

Assay Platform

Microplate reader

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Product details

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. The assay does not recognize NAD+/NADH. There is no need to purify NADP/NADPH from the sample mix. The enzyme cycling reaction significantly increases detection sensitivity. Results can be quantified using a plate reader at OD450nm.

NADP / NADPH assay protocol summary:
- extract samples from cells / tissues with extraction buffer and deproteinize with spin column
- for NADPH measurement, heat samples to 60°C for 30 min to decompose NAD+, cool on ice (this step not necessary for measurement of total NADP+/NADPH)
- add samples and standards to wells
- add reaction mix and incubate for 5 min at room temp to convert NADP to NADPH
- add NADPH developer and incubate for 1-4 hrs while reaction cycles
- analyze with microplate reader multiple times during the 1-4 hr incubation
- reaction can be stopped with stop solution

This product is manufactured by BioVision, an Abcam company and was previously called K347 NADP/NADPH Quantitation Colorimetric Kit. K347-100 is the same size as the 100 test size of ab65349.

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

The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.

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What's included?

{ "values": { "100Test": { "sellingSize": "100 Test", "publicAssetCode":"ab65349-100Test", "assetComponentDetails": [ { "size":"1 x 1.2 mL", "name":"Stop Solution II", "number":"AB65349-CMP05", "productcode":"" }, { "size":"1 x 1 Vial", "name":"NADPH Standard", "number":"AB65349-CMP01", "productcode":"" }, { "size":"1 x 200 µL", "name":"NADP Cycling Enzyme Mix", "number":"AB65349-CMP03", "productcode":"" }, { "size":"1 x 15 mL", "name":"NADP Cycling Buffer", "number":"AB65349-CMP02", "productcode":"AB323976" }, { "size":"1 x 50 mL", "name":"Extraction Buffer II", "number":"AB65349-CMP06", "productcode":"AB323943" }, { "size":"1 x 1 Vial", "name":"Developer Solution II", "number":"AB65349-CMP04", "productcode":"" } ] } } }
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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
Storage information
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

NADP/NADPH also known as Nicotinamide Adenine Dinucleotide Phosphate and its reduced form plays an essential role in cellular metabolism. It is a coenzyme with a molecular mass of around 743.4 g/mol. NADP/NADPH is ubiquitously expressed in cells due to its integral role in metabolic processes. It is found in high concentrations within the cytoplasm and chloroplasts where it functions in various biochemical reactions.
Biological function summary

NADPH facilitates reductive biosynthesis and maintains cellular redox status. It acts as a major electron donor in various anabolic reactions including lipid and nucleic acid synthesis. NADPH also contributes to the detoxification of reactive oxygen species and serves in the process of oxidative phosphorylation. Its cofactor activity associates it predominantly with enzymatic complexes such as the cytochrome P450 system.

Pathways

NADPH is fundamental in both the pentose phosphate pathway and the Calvin cycle. In the pentose phosphate pathway it provides reducing power important for fatty acid and cholesterol biosynthesis. In photosynthetic organisms NADPH functions in the Calvin cycle facilitating carbon fixation. The enzyme glucose-6-phosphate dehydrogenase related to NADPH production is integral in NADPH recycling.

Impaired NADPH function links to conditions like chronic granulomatous disease and diabetes. Chronic granulomatous disease occurs due to defects in the NADPH oxidase complex impairing the body’s ability to combat pathogens effectively. Diabetes has associations with oxidative stress influenced by imbalance in NADPH levels. In these diseases NADPH's interaction with proteins like superoxide dismutase affects oxidative damage management and overall immune response.
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Product protocols

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Target data

See full target information NADPH
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Publications (190)

Recent publications for all applications. Explore the full list and refine your search

Nature communications 16:5133 PubMed40461459

2025

ROMO1 overexpression protects the mitochondrial cysteinome from oxidations in aging.

Applications

Unspecified application

Species

Unspecified reactive species

Fengli Xu,Haipeng Huang,Kun Peng,Chongshu Jian,Hao Wu,Zhiwen Jing,Shan Qiu,Ying Chen,Keke Liu,Ling Fu,Yanru Wang,Jing Yang,Xiaotao Duan,Chu Wang,Heping Cheng,Xianhua Wang

Scientific reports 15:13635 PubMed40254646

2025

Disulfidptosis-related LncRNAs forecast the prognosis of acute myeloid leukemia.

Applications

Unspecified application

Species

Unspecified reactive species

Pei Xu,Xiaolin Sun,Lingxiao Pan,Jianfeng Zhu,Sixuan Qian

Frontiers in pharmacology 16:1563865 PubMed40166471

2025

Metformin upregulates circadian gene PER2 to inhibit growth and enhance the sensitivity of glioblastoma cell lines to radiotherapy via SIRT2/G6PD pathway.

Applications

Unspecified application

Species

Unspecified reactive species

Hailiang Li,Zheng Ma,Wanfu Yang,Yifan Zhang,Jinping Sun,Haifeng Jiang,Faxuan Wang,Li Hou,Hechun Xia

Cell communication and signaling : CCS 23:130 PubMed40065301

2025

SLC25A35 enhances fatty acid oxidation and mitochondrial biogenesis to promote the carcinogenesis and progression of hepatocellular carcinoma by upregulating PGC-1α.

Applications

Unspecified application

Species

Unspecified reactive species

Heng-Chao Yu,Lu Bai,Liang Jin,Yu-Jia Zhang,Zi-Han Xi,De-Sheng Wang

International journal of biological sciences 21:1275-1293 PubMed39897035

2025

Exosomal mediated lipid metabolism, ferritinophagy and CoQ-dependent pathway contributes to the ferroptosis of keratinocyte in SJS/TEN.

Applications

Unspecified application

Species

Unspecified reactive species

Chen Zhang,Pei Qiao,ChunYing Xiao,ZiPeng Cao,JiaoLing Chen,Hui Fang,JianKang Yang,ZeHua Kang,ErLe Dang,Shuai Shao,BingYu Pang,QingYang Li,ZhenLai Zhu,ShengXian Shen,Akito Hasegawa,Riichiro Abe,HongJiang Qiao,Gang Wang,Meng Fu

Immunology 174:450-461 PubMed39829079

2025

Mechanism Study of E2F8 Activation of SPC25-Mediated Glutamine Metabolism Promoting Immune Escape in Lung Adenocarcinoma.

Applications

Unspecified application

Species

Unspecified reactive species

Machang Luo,Lingyan Xie,Baoyan Lin,Xia Su,Rongzhang Liang,Zhiyi Ma,Youtang Li

Heliyon 10:e38718 PubMed39403483

2024

Mandatory role of endoplasmic reticulum in preserving NADPH regeneration in starved MDA-MB-231 breast cancer cells.

Applications

Unspecified application

Species

Unspecified reactive species

Sonia Carta,Vanessa Cossu,Francesca Vitale,Matteo Bauckneht,Maddalena Ghelardoni,Anna Maria Orengo,Serena Losacco,Daniela Gaglio,Silvia Bruno,Sabrina Chiesa,Silvia Ravera,Gianmario Sambuceti,Cecilia Marini

Marine drugs 22: PubMed39330302

2024

Fermented Fish Collagen Attenuates Melanogenesis via Decreasing UV-Induced Oxidative Stress.

Applications

Unspecified application

Species

Unspecified reactive species

Kyung-A Byun,So Young Lee,Seyeon Oh,Sosorburam Batsukh,Jong-Won Jang,Bae-Jin Lee,Kyoung-Min Rheu,Sichao Li,Min-Seok Jeong,Kuk Hui Son,Kyunghee Byun

Journal of biochemical and molecular toxicology 38:e23854 PubMed39287333

2024

Malate dehydrogenase-2 inhibition shields renal tubular epithelial cells from anoxia-reoxygenation injury by reducing reactive oxygen species.

Applications

Unspecified application

Species

Unspecified reactive species

Georgios Pissas,Maria Tziastoudi,Maria Divani,Christina Poulianiti,Maria Anna Polyzou Konsta,Evangelos Lykotsetas,Vasilios Liakopoulos,Ioannis Stefanidis,Theodoros Eleftheriadis

Respiratory research 25:321 PubMed39174953

2024

Vitamin D ameliorates particulate matter induced mitochondrial damages and calcium dyshomeostasis in BEAS-2B human bronchial epithelial cells.

Applications

Unspecified application

Species

Unspecified reactive species

Ju Chang-Chien,Jing-Long Huang,Hui-Ju Tsai,Shih-Ling Wang,Ming-Ling Kuo,Tsung-Chieh Yao
View all publications
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