Glyceraldehyde 3 Phosphate Dehydrogenase Activity Assay Kit (Colorimetric) (ab204732)
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
Overview
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Product name
Glyceraldehyde 3 Phosphate Dehydrogenase Activity Assay Kit (Colorimetric) -
Detection method
Colorimetric -
Sample type
Tissue, Adherent cells, Suspension cells -
Assay type
Enzyme activity -
Assay time
0h 20m -
Species reactivity
Reacts with: Mammals, Other species -
Product overview
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 -
Notes
GAPDH (Glyceraldehyde-3-Phosphate Dehydrogenase; EC 1.2.1.12) 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.
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Platform
Microplate reader
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components Identifier 100 tests GAPDH Assay Buffer WM 1 x 25ml GAPDH Developer Red 1 vial GAPDH Positive Control Orange 1 vial GAPDH Substrate Blue 1 x 200µl NADH Standard (500 nmol) Yellow 1 vial -
Research areas
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Function
Has 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. -
Pathway
Carbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 1/5. -
Sequence similarities
Belongs to the glyceraldehyde-3-phosphate dehydrogenase family. -
Post-translational
modificationsS-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus.
ISGylated. -
Cellular localization
Cytoplasm > 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
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Alternative names
- 38 kDa BFA-dependent ADP-ribosylation substrate
- aging associated gene 9 protein
- Aging-associated gene 9 protein
see all
Images
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Glyceraldehyde 3 Phosphate Dehydrogenase Activity Assay Kit (Colorimetric) (ab204732)
Typical NADH Standard Curve.
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Glyceraldehyde 3 Phosphate Dehydrogenase Activity Assay Kit (Colorimetric) (ab204732)GAPDH activity in the Positive Control, 3T3 cells lysate and MDA-MB-468 cells lysate.
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Glyceraldehyde 3 Phosphate Dehydrogenase Activity Assay Kit (Colorimetric) (ab204732)GAPDH specific activity calculated from 3T3 cell lysate (11.1 µg protein), and MDA-MB-468 cell lysate (2.76 µg protein).