Key features and details
- Assay type: Enzyme activity
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 20 min
- Sample type: Adherent cells, Suspension cells, Tissue
Product nameGlyceraldehyde 3 Phosphate Dehydrogenase Activity Assay Kit (Colorimetric)
See all GAPDH kits
Sample typeTissue, Adherent cells, Suspension cells
Assay typeEnzyme activity
Assay time0h 20m
Species reactivityReacts with: Mammals, Other species
Glyceraldehyde 3 Phosphate Dehydrogenase Activity Assay Kit (Colorimetric) (ab204732) provides a simple and sensitive method for monitoring GAPDH activity in various samples. In this assay, GAPDH catalyzes conversion of GAP into BPG and an intermediate, which reacts with a developer to form a colored product that absorbs maximally at 450 nm. Our high-throughput adaptable assay can detect GAPDH activity as low as 100 µU/mL in a variety of samples.
GAPDH assay protocol summary:
- add samples and standards to wells
- add reaction mix
- analyze with microplate reader for 10 min every 2-3 min
This product is manufactured by BioVision, an Abcam company and was previously called K680 GAPDH Activity Assay Kit. K680-100 is the same size as the 100 test size of ab204732.
GAPDH (Glyceraldehyde-3-Phosphate Dehydrogenase; EC 22.214.171.124) catalyzes the conversion of Glyceraldehyde-3-Phosphate (GAP) to 1, 3-Bisphosphate Glycerate (BPG) and plays a key role in glycolysis. The enzyme is involved in cellular processes such as apoptosis, membrane trafficking, iron metabolism and nuclear translocation. GAPDH (housekeeping gene) expression is stable and constitutive. Deregulation of GAPDH activity is associated with abnormal cell proliferation and carcinogenesis. Accurate quantitation of GAPDH activity is important for diagnosing diseases and studying normal cellular physiology.
Storage instructionsStore at -20°C. Please refer to protocols.
Components 100 tests GAPDH Assay Buffer 1 x 25ml GAPDH Developer 1 vial GAPDH Positive Control 1 vial GAPDH Substrate 1 x 200µl NADH Standard (500 nmol) 1 vial
- Pathways and Processes
- Metabolic signaling pathways
- Energy transfer pathways
- Energy Metabolism
FunctionHas both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis. Nuclear functions are probably due to the nitrosylase activity that mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2 and PRKDC (By similarity). Glyceraldehyde-3-phosphate dehydrogenase is a key enzyme in glycolysis that catalyzes the first step of the pathway by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate.
PathwayCarbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 1/5.
Sequence similaritiesBelongs to the glyceraldehyde-3-phosphate dehydrogenase family.
modificationsS-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus.
Cellular localizationCytoplasm > cytosol. Nucleus. Cytoplasm > perinuclear region. Membrane. Translocates to the nucleus following S-nitrosylation and interaction with SIAH1, which contains a nuclear localization signal (By similarity). Postnuclear and Perinuclear regions.
- Information by UniProt
- 38 kDa BFA-dependent ADP-ribosylation substrate
- aging associated gene 9 protein
- Aging-associated gene 9 protein
ab204732 has been referenced in 33 publications.
- Jin X et al. Glucose-6-phosphate dehydrogenase exerts antistress effects independently of its enzymatic activity. J Biol Chem 298:102587 (2022). PubMed: 36243112
- Petruzzelli M et al. Early Neutrophilia Marked by Aerobic Glycolysis Sustains Host Metabolism and Delays Cancer Cachexia. Cancers (Basel) 14:N/A (2022). PubMed: 35205709
- Rashed FB et al. Cellular mechanism of action of 2-nitroimidazoles as hypoxia-selective therapeutic agents. Redox Biol 52:102300 (2022). PubMed: 35430547
- Haythorne E et al. Altered glycolysis triggers impaired mitochondrial metabolism and mTORC1 activation in diabetic β-cells. Nat Commun 13:6754 (2022). PubMed: 36376280
- Liu X et al. Glyceraldehyde-3-Phosphate Dehydrogenase Restricted in Cytoplasmic Location by Viral GP5 Facilitates Porcine Reproductive and Respiratory Syndrome Virus Replication via Its Glycolytic Activity. J Virol 95:e0021021 (2021). PubMed: 34160254