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MAB374 Sigma-Aldrich

Anti-Glyceraldehyde-3-Phosphate Dehydrogenase Antibody, clone 6C5

Synonym: glyceraldehyde 3-phosphate dehydrogenase, aging-associated gene 9 protein, glyceraldehyde-3-phosphate dehydrogenase, GAPDH, G3PDH, GAPD, OK/SW-cl.12, MGC88685, G3PD, EC 1.2.1.12

  •  eCl@ss 32160702

  •  NACRES NA.41

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Properties

Related Categories Alphabetical Index, Antibodies, GD-GL, Primary Antibodies
clone   6C5, monoclonal
biological source   mouse
application(s)   ELISA: suitable
  immunocytochemistry: suitable
  immunofluorescence: suitable
  immunohistochemistry: suitable
  immunoprecipitation (IP): suitable
  western blot: suitable
species reactivity   rabbit, feline, mouse, rat, fish, pig, canine, human
should not react with   E. coli
shipped in   wet ice
isotype   IgG1
antibody product type   primary antibodies
mfr. no.   Chemicon®
NCBI accession no.   NP_002037
UniProt accession no.   P04406

Description

General description

Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is an ubiquitous glycolytic enzyme present in reasonably high levels in almost all tissues. As a ′house-keeping′ enzyme, it catalyzes the synthesis of 1,3-biphosphoglycerate, a "high energy" intermediate used for the synthesis of ATP. Besides its cytoplasmic action in metabolism it is also involved in the initial stages of apoptosis or oxidative stress response where GAPDH is translocated to the nucleus. Such actions may reflect the role of GAPDH in DNA repair or as one nuclear carrier for apoptotic molecules. GAPDH has also been found to bind specifically to proteins implicated in the pathogenesis of a variety of neurodegenerative disorders including the beta-amyloid precursor protein and the huntingtin protein where decreased function of GAPDH is associated with Alzheimer′s and Huntington′s disease fibroblasts. GAPDH has also been identified as a potential target for nitric oxide (NO)-mediated cellular toxicity. The complete and functional enzyme is a tetramer with each of four identical subunits occupying the vertex of a tetrahedron. Binding domains also include one principally interacting with NAD+ and another interacting with glyceraldehyde 3-phosphate (GAP).

Specificity

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from skeletal muscle. Antibody also recognizes cardiac GAPDH. GAPDH enzyme is detected in many non-muscle cells lines including HeLa, HCT-116 cells, U937 and THP-1 cells among others.

Human, porcine, canine, rat, mouse, rabbit, cat, and fish skeletal muscle. It has been reported that this antibody does not react with GAPDH from E. coli.

Immunogen

Glyceraldehyde-3-phosphate dehydrogenase from rabbit muscle.

Application

ELISA:
1:1,000,000 dilution of a previous lot ws used for Immunoaffinity purification of GAPDH.

Immunocytochemistry:
1:100 to 1:300 dilution of a previous lot was used in immunocytochemistry in PBS-BSA 10 mg/mL. 4% PFA, with 0.02% PBS-Triton X-100; 3 min RT (Griffoni, 2001).

Western blot:
1:100 to 1:300. Recognizes a 36kDa band of the reduced monomer. Non-reduced GAPDH runs as a 146kDa tetramer.

Immunoaffinity purification of GAPDH.

Optimal working dilutions must be determined by end user.

Research Category
Metabolism

Research Sub Category
Enzymes & Biochemistry

Target description

38 kDa

Physical form

Format: Purified

Protein A Purfied

Purified Mouse monoclonal IgG1 in PBS, pH 7.4 with 0.1% sodium azide.

Storage and Stability

Maintain at 2-8ºC in undiluted aliquots for up to 6 months after date of receipt.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Quality

Evaluated by Western Blot on A431 lysates.

Western Blot Analysis:
1:500 dilution of this antibody detected GLYCERALDEHYDE-3-PDH on 10 μg of A431 lysates.

Analysis Note

Control
It is expressed in all cells.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Safety & Documentation

Safety Information

Safety Information for this product is unavailable at this time.
Protocols & Articles

Articles

Antibody Basics

Immunoglobulins (Igs) are produced by B lymphocytes and secreted into plasma. The Ig molecule in monomeric form is a glycoprotein with a molecular weight of approximately 150 kDa that is shaped more ...
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Protocols

Western Blot Protocol | Immunoblotting Protocol

Western Blotting refers to the electrophoretic transfer of proteins from sodium dodecyl sulfate polyacrylamide gels to sheets of PVDF or nitrocellullose membrane, followed by immunodetection of prote...
Keywords: AGE, Buffers, Cell disruption, Detection methods, Detergents, Dialysis, Electroblotting, Electrophoresis, Enzyme activity, Gel electrophoresis, Immunoprecipitation, PAGE, Protein extraction, Purification, Sample preparations, Western blot

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Peer-Reviewed Papers
15

References

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