AB176723

NAD/NADH Assay Kit (Fluorometric)

Name: NAD/NADH Assay Kit. Fluorometric. Quantitative. (ab176723) | Abcam
Brand: Abcam Limited
SKU: AB176723
Rating: 3 (2 reviews)

(2 Reviews)

(19 Publications)

NAD/NADH Assay Kit (Fluorometric) ab176723 provides a convenient method for sensitive detection of NAD, NADH and their ratio. Readout on any fluorometric (Ex/Em 540/590 nm) plate reader.

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Functional Studies - NAD/NADH Assay Kit (Fluorometric) (AB176723)

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Supplier Data

Functional Studies - NAD/NADH Assay Kit (Fluorometric) (AB176723)

Total NADH and NAD Dose Response Curve.

Total NADH and NAD, and their extract dose response were measured with NAD/NADH Assay Kit (Fluorometric) (ab176723) in a 96-well black plate using a Gemini microplate reader (Molecular Devices). The signal was acquired at Ex/Em = 540/590 nm (cut off at 570 nm) 30 minutes after adding NAD/NADHH reaction mixture. The blank signal was subtracted from the values for those wells with the NADH reactions.

Key facts

Detection method

Colorimetric/Fluorometric

Sample types

Tissue Lysate, Cell Lysate

Results type

Quantitative

Assay time

2h 30m

Assay Platform

Microplate reader

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Product details

NAD/NADH Assay Kit (Fluorometric) ab176723 provides a convenient method for sensitive detection of NAD, NADH and their ratio.

The enzymes used in the NAD/NADH assay protocol specifically recognize NAD/NADH in an enzyme cycling reaction that significantly increases detection sensitivity. In addition, this assay has very low background since it is run in the red visible range that considerably reduces the interference from biological samples.

There is no need to purify NAD/NADH from sample mix. The assay can be performed in a convenient 96-well or 384-well microtiter-plate format.

The NAD/NADH assay signal can be easily read by either a fluorescence microplate reader at Ex/Em 530 - 570/590 - 600 nm (max Ex/Em 540/590 nm) or an absorbance microplate reader at ~576 nm.

This kit provides NAD and NADH extraction buffer, and cell lysis buffer for your convenience. It has been frequently used for determining NAD/NADH from cell lysates.

NAD/NADH assay protocol summary:
- add standards and samples for NAD, NADH, total NAD/NADH measurement to wells
- add NADH extraction solution to NADH wells, incubate for 10-15 min, and add NAD extraction solution to neutralize
- add NAD extraction solution to NAD wells, incubate for 10-15 min, and add NADH extraction solution to neutralize
- add NAD/NADH control solution to standard and total NAD/NADH wells, incubate for 10-15 min, and add NAD/NADH control solution
- add NADH reaction mix and incubate for 30 min to 2 hr
- analyze with a microplate reader

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What's included?

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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.

NAD/NADH often called nicotinamide adenine dinucleotide acts as an important coenzyme in redox reactions. It has a molecular mass of 663.43 g/mol. NAD/NADH exists widely across all living cells functioning predominantly in the cytoplasm and mitochondria. It is an important component in metabolic pathways serving as an electron carrier in processes that generate and use ATP. This coenzyme oscillates between oxidized (NAD⁺) and reduced (NADH) forms essential for energy production.

Biological function summary

The oscillation between NAD⁺ and NADH enables cells to maintain redox homeostasis. It is not part of a larger protein complex but plays an important role in several biological reactions by transferring electrons. NAD⁺ works as an electron acceptor while NADH serves as an electron donor. Their balance influences various cellular processes including DNA repair and gene expression regulation. This coenzyme is essential to the enzymatic activity of dehydrogenases which are pivotal for the energy metabolism.

Pathways

NAD+/NADH balance is vital in glycolysis and the citric acid cycle. Glycolysis uses NAD⁺ to help break down glucose into pyruvate while the citric acid cycle further processes acetyl-CoA producing NADH. NADH produced in these pathways then enters the electron transport chain driving ATP synthesis. This coenzyme also connects with sirtuins a family of proteins known for regulating cellular health and longevity through NAD⁺-dependent deacetylase activity.

NAD/NADH imbalance links with metabolic conditions like diabetes and neurodegenerative diseases such as Alzheimer's. In diabetes altered NAD+/NADH ratio can impact insulin secretion and glucose metabolism. Alzheimer's disease associates with disrupted NAD+ levels that affect sirtuins' function leading to impaired cellular repair mechanisms. These connections illustrate the coenzyme's significant effect on human health and highlight its potential as a therapeutic target.

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Product protocols

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Target data

See full target information Disodium nicotinamide adenine dinucleotide

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Publications (19)

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

Cell death & disease 16:489 PubMed40610445

2025

The dual mechanism of mA demethylase ALKBH5 in regulating energy metabolism during exposure to MC-LR.

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Xiaoya Sun,Qinmei Tan,Yue Yang,Jia Wei,Xiaodie Zhou,Shanshan Gao,Fei Yang

Redox biology 85:103730 PubMed40554101

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Endothelial soluble epoxide hydrolase links polyunsaturated fatty acid metabolism to oxidative stress and atherosclerosis progression.

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Minghua Cao,Mengqing Li,Xixuan Li,Yuanyuan Li,Yunyun Chen,Maria-Kyriaki Drekolia,Xiang Cheng,Fredy Delgado Lagos,Sofia-Iris Bibli,Jiong Hu

Redox biology 69:103014 PubMed38171255

2024

Loss of cardiac mitochondrial complex I persulfidation impairs NAD homeostasis in aging.

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Maria-Kyriaki Drekolia,Christina Karantanou,Ilka Wittig,Yuanyuan Li,Dominik C Fuhrmann,Bernhard Brüne,Antonia Katsouda,Jiong Hu,Andreas Papapetropoulos,Sofia-Iris Bibli

Nature communications 14:7674 PubMed37996418

2023

A reversible state of hypometabolism in a human cellular model of sporadic Parkinson's disease.

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Sebastian Schmidt,Constantin Stautner,Duc Tung Vu,Alexander Heinz,Martin Regensburger,Ozge Karayel,Dietrich Trümbach,Anna Artati,Sabine Kaltenhäuser,Mohamed Zakaria Nassef,Sina Hembach,Letyfee Steinert,Beate Winner,Winkler Jürgen,Martin Jastroch,Malte D Luecken,Fabian J Theis,Gil Gregor Westmeyer,Jerzy Adamski,Matthias Mann,Karsten Hiller,Florian Giesert,Daniela M Vogt Weisenhorn,Wolfgang Wurst

Redox biology 67:102943 PubMed37883843

2023

Adaptive responses of neuronal cells to chronic endoplasmic reticulum (ER) stress.

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Thu Nguyen Minh Pham,Natarajan Perumal,Caroline Manicam,Marion Basoglu,Stefan Eimer,Dominik C Fuhrmann,Claus U Pietrzik,Albrecht M Clement,Hagen Körschgen,Jana Schepers,Christian Behl

Molecular cancer research : MCR 22:197-208 PubMed37878010

2023

Mitochondrial Translocase TOMM22 Is Overexpressed in Pancreatic Cancer and Promotes Aggressive Growth by Modulating Mitochondrial Protein Import and Function.

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Mary Oluwadamilola Haastrup,Kunwar Somesh Vikramdeo,Shashi Anand,Mohammad Aslam Khan,James Elliot Carter,Seema Singh,Ajay Pratap Singh,Santanu Dasgupta

Cells 12: PubMed37408187

2023

Live Cell Imaging of ATP Levels Reveals Metabolic Compartmentalization within Motoneurons and Early Metabolic Changes in ALS Motoneurons.

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Vitaly L Zimyanin,Anna-Maria Pielka,Hannes Glaß,Julia Japtok,Dajana Großmann,Melanie Martin,Andreas Deussen,Barbara Szewczyk,Chris Deppmann,Eli Zunder,Peter M Andersen,Tobias M Boeckers,Jared Sterneckert,Stefanie Redemann,Alexander Storch,Andreas Hermann

Nature 617:386-394 PubMed37100912

2023

A druggable copper-signalling pathway that drives inflammation.

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Stéphanie Solier,Sebastian Müller,Tatiana Cañeque,Antoine Versini,Arnaud Mansart,Fabien Sindikubwabo,Leeroy Baron,Laila Emam,Pierre Gestraud,G Dan Pantoș,Vincent Gandon,Christine Gaillet,Ting-Di Wu,Florent Dingli,Damarys Loew,Sylvain Baulande,Sylvère Durand,Valentin Sencio,Cyril Robil,François Trottein,David Péricat,Emmanuelle Näser,Céline Cougoule,Etienne Meunier,Anne-Laure Bègue,Hélène Salmon,Nicolas Manel,Alain Puisieux,Sarah Watson,Mark A Dawson,Nicolas Servant,Guido Kroemer,Djillali Annane,Raphaël Rodriguez

Frontiers in bioengineering and biotechnology 10:971932 PubMed36561039

2022

Modulation of sirtuins during monolayer chondrocyte culture influences cartilage regeneration upon transfer to a 3D culture environment.

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Hannah K Heywood,Stephen D Thorpe,Renos M Jeropoulos,Paul W Caton,David A Lee

Biomolecules 12: PubMed36551225

2022

Effects of Calcium Ions on the Antimicrobial Activity of Gramicidin A.

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Shang-Ting Fang,Shu-Hsiang Huang,Chin-Hao Yang,Jen-Wen Liou,Hemalatha Mani,Yi-Cheng Chen

View all publications

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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NAD/NADH Assay Kit (Fluorometric)

Functional Studies - NAD/NADH Assay Kit (Fluorometric) (AB176723)

Key facts

Detection method

Sample types

Results type

Assay time

Assay Platform

Complementary Products

Assay Kits

Secondary Antibodies

Assay Kits

Primary Antibodies

Product details

What's included?

Properties and storage information

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Publications (19)

Product promise