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G9545

Sigma-Aldrich

Anti-GAPDH antibody produced in rabbit

~1 mg/mL, affinity isolated antibody, buffered aqueous solution

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Synonym(s):
Anti Gapdh, Anti Gapdh Antibody, Anti-Gapdh, GAPDH Antibody - Anti-GAPDH antibody produced in rabbit, Gapdh Antibody, Gapdh Antibody Sigma, Anti-38-kD component, Anti-G3PD, Anti-GAPD, Anti-Glyceraldehyde-3-phosphate dehydrogenase, Anti-OCAS, Anti-OCT1 coactivator in S phase, Anti-p38 component
MDL number:
NACRES:
NA.41

biological source

rabbit

Quality Level

conjugate

unconjugated

antibody form

affinity isolated antibody

antibody product type

primary antibodies

clone

polyclonal

form

buffered aqueous solution

mol wt

antigen ~36 kDa

species reactivity

mouse, rat, human

concentration

~1 mg/mL

technique(s)

immunoprecipitation (IP): 5-10 μg using mouse NIH3T3 cell lysates
indirect immunofluorescence: 5-10 μg/mL using rat NRK cells
western blot: 0.1-0.2 μg/mL using whole extract of human HeLa cells

UniProt accession no.

shipped in

dry ice

storage temp.

−20°C

target post-translational modification

unmodified

Gene Information

human ... GAPDH(2597)
mouse ... Gapdh(14433)
rat ... Gapdh(24383)

General description

The GAPDH (glyceraldehyde-3-phosphate dehydrogenase) gene is mapped to human chromosome 12p13.3 and encodes a tetramer containing identical chains. The enzyme is commonly known as glycolytic enzyme and contains a binding site for NAD+(nicotinamide adenine diphosphate) and glyceraldehyde-3-phosphate. GAPDH contributes to 10-20% of the total cellular protein and is considered to be evolutionarily conserved.

Specificity

Anti-GAPDH recognizes human, mouse, and rat GAPDH.

Immunogen

Synthetic peptide corresponding to amino acids of mouse GAPDH, conjugated to KLH via an N-terminal cysteine residue. The corresponding sequence is identical in rat and differs by two amino acids in humans.

Application

Anti-GAPDH antibody produced in rabbit is suitable as a primary antibody for western blotting using:
  • human cancer cell extract
  • extract from muscle tissue from old mice exhibiting sarcopenia
  • trabeculae or samples of the mice heart myocardium
  • mouse embryonic fibroblasts
It is suitable for immunoprecipitation using mouse NIH3T3 cell lysates with 5-10μg, for indirect immunofluorescence using rat NRK cells at a working concentration of 5-10μg/mL. It is also suitable for western blotting at a working concentration of 0.1-0.2μg/mL using a whole extract of human HeLa cells.

Biochem/physiol Actions

The enzyme glyceraldehyde-3-phosphate dehydrogenase catalyzes the reversible oxidative phosphorylation of glyceraldehyde-phosphate in the presence of inorganic phosphate and NAD, 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. 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. In response to oxidative stress, GAPDH induces apoptosis.

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 “frost free” freezers, is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilutions 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.

Storage Class Code

10 - Combustible liquids

WGK

WGK 1

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Regulatory Information

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Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) aggregation causes mitochondrial dysfunction during oxidative stress-induced cell death.
Nakajima H, et al.
The Journal of Biological Chemistry, M116-M116 (2017)
Jeffrey G Dickhout et al.
The Journal of biological chemistry, 287(10), 7603-7614 (2012-01-05)
The integrated stress response mediated by eukaryotic translation initiation factor 2α (eIF2α) phosphorylation maintains cellular homeostasis under endoplasmic reticulum (ER) stress. eIF2α phosphorylation induces activating transcription factor 4 (ATF4), a basic leucine zipper transcription factor that regulates the expression of
Role of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in DNA repair
Kosova A A, et al.
Biochemistry (Moscow), 82(6), 643-654 (2017)
Abiodun T Kukoyi et al.
American journal of physiology. Cell physiology, 317(2), C390-C397 (2019-05-16)
Chronic HIV infection causes redox stress and increases the risk of acute and chronic lung injury, even when individuals are adherent to antiretroviral therapy. HIV-1 transgene expression in rats inhibits nuclear factor (erythroid-derived 2)-like 2 (Nrf2), which regulates antioxidant defenses
Robbe Naert et al.
International journal of molecular sciences, 20(14) (2019-07-13)
The increase in cytosolic Ca2+ is essential in key effector functions of dendritic cells (DCs), including differentiation, maturation, cytokine expression, and phagocytosis. Although several Ca2+-permeable ion channels have been described in DCs, the contribution of transient receptor potential (TRP) channels

Articles

The field of proteomics is continually looking for new ways to investigate protein dynamics within complex biological samples. Recently, many researchers have begun to use RNA interference (RNAi) as a method of manipulating protein levels within their samples, but the ability to accurately determine these protein amounts remains a challenge. Fortunately, over the past decade, the field of proteomics has witnessed significant advances in the area of mass spectrometry. These advances, both in instrumentation and methodology, are providing researchers with sensitive assays for both identification and quantification of proteins within complex samples. This discussion will highlight some of these methodologies, namely the use of Multiple Reaction Monitoring (MRM) and Protein-AQUA.

Loading controls in western blotting application.

We presents an article about the Warburg effect, and how it 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 cells show an increased dependence on glycolysis to meet their energy needs, regardless of whether they were well-oxygenated or not.

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