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G8795 Sigma-Aldrich

Anti-GAPDH antibody, Mouse monoclonal

clone GAPDH-71.1, purified from hybridoma cell culture

Synonym: Anti-Glyceraldehyde-3-phosphate dehydrogenase, Anti-G3PD, Anti-G3PDH, GAPDH Antibody - Monoclonal Anti-GAPDH antibody produced in mouse, Gapdh Antibody, Loading Control

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Properties

Related Categories Aerobic Glycolysis (the Warburg Effect), Alphabetical Index, Antibodies, Antibodies Against Proteins Involved in Glucose Metabolism, Cancer Metabolism,
conjugate   unconjugated
clone   GAPDH-71.1, monoclonal
biological source   mouse
application(s)   immunocytochemistry: suitable
  indirect ELISA: suitable
  microarray: suitable
  western blot: 0.025-0.05 μg/mL using A431 total cell extract
species reactivity   mouse, mink, rabbit, rat, human, hamster, canine, turkey, chicken, monkey, bovine
should not react with   prokaryotes
mol wt   antigen ~37 kDa
form   buffered aqueous solution
shipped in   dry ice
storage temp.   −20°C
antibody form   purified from hybridoma cell culture
  purified immunoglobulin
isotype   IgM
antibody product type   primary antibodies
concentration   ~1 mg/mL
Featured Industry   Research Pathology
UniProt accession no.   P04406
Gene Information   human ... GAPDH(2597)
mouse ... Gapdh(14433)
rat ... Gapdh(24383)

Description

General description

Monoclonal Anti-GAPDH (mouse IgM isotype) is derived from the hybridoma GAPDH-71.1 produced by the fusion of mouse myeloma cells (NS1 cells) and splenocytes from BALB/c mice immunized with rabbit GAPDH (glyceraldehyde-3-phosphate dehydrogenase). The gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is mapped to human chromosome 12p13. The protein localizes in the cytoplasm but can be translocated to the nucleus depending on cellular conditions.

Specificity

Monoclonal Anti-GAPDH recognizes human, monkey, bovine, canine, rat, mouse, hamster, mink, rabbit, chicken, and turkey GAPDH. It does not cross-react with non-vertebrate and prokaryotic species.

Immunogen

Rabbit GAPDH.

Application

Monoclonal Anti-GAPDH antibody produced in mouse is suitable for western blotting using:
• protein extracted from heart tissue of mice at a working dilution of 1:25,000
• myelin and axogliasomal fractions from human CNS
• nuclear and cytoplasmic fractions from TBP-13Q and TBP-105Q PC12 cells following recovery from heat shock
• protein from bovine immortalized luteal endothelial cells
• renal tubular epithelial cell extract
• proteins from mouse embryonic fibroblasts
• protein extract from ventricular myocardium tissues
• A431 total cell extract at a working concentration of 0.025-0.05μg/mL
It is also suitable for immunostaining using leiomyomas and leiomyosarcomas. The antibody can also be used for immunocytochemistry, indirect ELISA and microarray.

Physical form

Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 15 mM sodium azide.

Storage and Stability

For continuous use, store at 2-8 °C for up to one month.
For extended storage, freeze in working aliquots. Repeated freezing and thawing, or storage in “frostfree” freezers, is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilution samples should be discarded if not used within 12 hours.

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.

Biochem/physiol Actions

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a tetramer containing identical chains. It catalyzes the reversible oxidative phosphorylation of glyceraldehyde-phosphate, which is a critical energy-yielding step in carbohydrate metabolism. It binds to several proteins including actin, tubulin, amyloid precursor, polyglutamine peptides, DRPLA (dentatorubral-pallidoluysian atrophy), and huntingtin. Protein kinase Cι/λbinds and phosphorylates GAPDH. Phosphorylated GAPDH associates with cytoskeletal elements and controls microtubule dynamics in the early secretory pathway. Poly(ADP-ribose) polymerase-1 (PARP1) interacts with GAPDH and thereby mediates brain damage in the presence of oxidative/nitrosative stress. GAPDH forms a part of OCA-S, the multicomponent OCT1 (octamer-motif-binding factor) coactivator complex, which is involved in the S phase-dependent histone H2B transcription. This association is responsible for linking H2B transcriptional machinery to cell cycle regulation and to the cellular metabolic state. GAPDH is also a component of the functional GAIT (interferon-γ-activated inhibitor of translation) mRNP (messenger ribonucleoprotein). GAPDH expression is dysregulated during melanoma progression.

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
3

Documents

Certificate of Analysis

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Certificate of Origin

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Protocols & Articles

Articles

Aerobic Glycolysis and the Warburg Effect

The Warburg effect is the enhanced conversion of glucose to lactate observed in tumor cells, even in the presence of normal levels of oxygen. Otto Heinrich Warburg demonstrated in 1924 that cancer ce...
Keywords: Aerobic, Cancer, Cell proliferation, Clinical, Glycolysis, PAGE, Phosphorylations, Transcription

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, 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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