Product: GAPDH Antibody
Catalog: T0004
Description: Mouse monoclonal antibody to GAPDH
Application: WB ELISA
Reactivity: Human, Mouse, Rat, Pig, Bovine, Sheep, Rabbit, Goat, Guinea pig, Dog, Monkey, Hamster, Chicken, Plants, Rice, Fish
Mol.Wt.: 34KD; 36kD(Calculated).
Uniprot: P04406
RRID: AB_2833041

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Product Info

Source:
Mouse
Application:
WB 1:3000-1:10000
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat,Pig,Bovine,Sheep,Rabbit,Goat,Guinea pig,Dog,Monkey,Hamster,Chicken,Plants,Rice,Fish
Clonality:
Monoclonal [2F40]
Specificity:
GAPDH Mouse Monoclonal antibody detects endogenous levels of total GAPDH protein.
RRID:
AB_2833041
Cite Format: Affinity Biosciences Cat# T0004, RRID:AB_2833041.
Conjugate:
Unconjugated.
Purification:
Affinity-chromatography.
Storage:
Mouse IgG1 in phosphate buffered saline (without Mg2+ and Ca2+), pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

GAPDH; A1 40 kd subunit; Activator 1 40 kd subunit; G3PD; GAPD; G3pdh; Rfc40; Rf-c 40 kd subunit;

Immunogens

Immunogen:

Full-length GAPDH protein of human.

Uniprot:
Gene(ID):
Description:
Glyceraldehyde 3 phosphate dehydrogenase (GAPDH) is well known as one of the key enzymes involved in glycolysis. GAPDH is constitutively abundant expressed in almost cell types at high levels, therefore antibodies against GAPDH are useful as loading controls for Western Blotting. Some pathology factors, such as hypoxia and diabetes, increased or decreased GAPDH expression in certain cell types.
Sequence:
MGKVKVGVNGFGRIGRLVTRAAFNSGKVDIVAINDPFIDLNYMVYMFQYDSTHGKFHGTVKAENGKLVINGNPITIFQERDPSKIKWGDAGAEYVVESTGVFTTMEKAGAHLQGGAKRVIISAPSADAPMFVMGVNHEKYDNSLKIISNASCTTNCLAPLAKVIHDNFGIVEGLMTTVHAITATQKTVDGPSGKLWRDGRGALQNIIPASTGAAKAVGKVIPELNGKLTGMAFRVPTANVSVVDLTCRLEKPAKYDDIKKVVKQASEGPLKGILGYTEHQVVSSDFNSDTHSSTFDAGAGIALNDHFVKLISWYDNEFGYSNRVVDLMAHMASKE

Research Backgrounds

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. Modulates the organization and assembly of the cytoskeleton. Facilitates the CHP1-dependent microtubule and membrane associations through its ability to stimulate the binding of CHP1 to microtubules (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. Component of the GAIT (gamma interferon-activated inhibitor of translation) complex which mediates interferon-gamma-induced transcript-selective translation inhibition in inflammation processes. Upon interferon-gamma treatment assembles into the GAIT complex which binds to stem loop-containing GAIT elements in the 3'-UTR of diverse inflammatory mRNAs (such as ceruplasmin) and suppresses their translation.

PTMs:

S-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus (By similarity). S-nitrosylation of Cys-247 is induced by interferon-gamma and LDL(ox) implicating the iNOS-S100A8/9 transnitrosylase complex and seems to prevent interaction with phosphorylated RPL13A and to interfere with GAIT complex activity.

ISGylated.

Sulfhydration at Cys-152 increases catalytic activity.

Oxidative stress can promote the formation of high molecular weight disulfide-linked GAPDH aggregates, through a process called nucleocytoplasmic coagulation. Such aggregates can be observed in vivo in the affected tissues of patients with Alzheimer disease or alcoholic liver cirrhosis, or in cell cultures during necrosis. Oxidation at Met-46 may play a pivotal role in the formation of these insoluble structures. This modification has been detected in vitro following treatment with free radical donor (+/-)-(E)-4-ethyl-2-[(E)-hydroxyimino]-5-nitro-3-hexenamide. It has been proposed to destabilize nearby residues, increasing the likelihood of secondary oxidative damages, including oxidation of Tyr-45 and Met-105. This cascade of oxidations may augment GAPDH misfolding, leading to intermolecular disulfide cross-linking and aggregation.

Succination of Cys-152 and Cys-247 by the Krebs cycle intermediate fumarate, which leads to S-(2-succinyl)cysteine residues, inhibits glyceraldehyde-3-phosphate dehydrogenase activity. Fumarate concentration as well as succination of cysteine residues in GAPDH is significantly increased in muscle of diabetic mammals. It was proposed that the S-(2-succinyl)cysteine chemical modification may be a useful biomarker of mitochondrial and oxidative stress in diabetes and that succination of GAPDH and other thiol proteins by fumarate may contribute to the metabolic changes underlying the development of diabetes complications.

Subcellular Location:

Cytoplasm>Cytosol. Nucleus. Cytoplasm>Perinuclear region. Membrane. Cytoplasm>Cytoskeleton.
Note: Translocates to the nucleus following S-nitrosylation and interaction with SIAH1, which contains a nuclear localization signal (By similarity). Postnuclear and Perinuclear regions.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Subunit Structure:

Homotetramer. Interacts with TPPP; the interaction is direct. Interacts (when S-nitrosylated) with SIAH1; leading to nuclear translocation. Interacts with RILPL1/GOSPEL, leading to prevent the interaction between GAPDH and SIAH1 and prevent nuclear translocation. Interacts with CHP1; the interaction increases the binding of CHP1 with microtubules. Associates with microtubules (By similarity). Interacts with EIF1AD, USP25, PRKCI and WARS1. Interacts with phosphorylated RPL13A; inhibited by oxidatively-modified low-densitity lipoprotein (LDL(ox)). Component of the GAIT complex. Interacts with FKBP6; leading to inhibit GAPDH catalytic activity.

Family&Domains:

The [IL]-x-C-x-x-[DE] motif is a proposed target motif for cysteine S-nitrosylation mediated by the iNOS-S100A8/A9 transnitrosylase complex.

Belongs to the glyceraldehyde-3-phosphate dehydrogenase family.

Research Fields

· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway.   (View pathway)

· Human Diseases > Neurodegenerative diseases > Alzheimer's disease.

· Metabolism > Carbohydrate metabolism > Glycolysis / Gluconeogenesis.

· Metabolism > Global and overview maps > Metabolic pathways.

· Metabolism > Global and overview maps > Carbon metabolism.

· Metabolism > Global and overview maps > Biosynthesis of amino acids.

References

1). Lipopolysaccharide-binding protein expression is increased by stress and inhibits monoamine synthesis to promote depressive symptoms. Immunity, 2023 (PubMed: 36854305) [IF=32.4]

2). Sulforaphane elicts dual therapeutic effects on Renal Inflammatory Injury and crystal deposition in Calcium Oxalate Nephrocalcinosis. Theranostics, 2020 (PubMed: 32641994) [IF=12.4]

Application: WB    Species: mice    Sample: BMDMs

Figure 3. Nrf2 suppresses TLR4 and IRF1 levels and promotes M2Mϕ polarization in vitro. (A) BMDM and COM-TECs coculture schematic diagram. Western blot (B) and qPCR (C) analyses of Nrf2, TLR4, IRF1, iNOS, and ARG-1 levels in BMDMs co-cultured with increasing COM dose stimulated TECs. GAPDH was used as an internal control. (D, F) Western blot detection of Nrf2, TLR4, IRF1, iNOS, and ARG-1 levels following SFN treatment or Nrf2 upregulation/downregulation in BMDMs co-cultured with COM-stimulated TECs. GAPDH was used as an internal control. (E, G) The distribution of iNOS (green) and ARG-1 (red) in BMDMs according to immunofluorescence. (H, I) Flow cytometric analysis of the polarization state of BMDMs using anti-CD11c and anti-CD206 in F4/80+ and CD11b+ cells. The data are shown as the mean ± standard error (SE) of three independent experiments. *P < 0.05; **P < 0.01, as determined by Student’s t test (C) or one-way ANOVA (E, G, H, I).

3). Stearoyl-CoA desaturase-1 promotes colorectal cancer metastasis in response to glucose by suppressing PTEN. JOURNAL OF EXPERIMENTAL & CLINICAL CANCER RESEARCH, 2018 (PubMed: 29530061) [IF=11.3]

Application: WB    Species: human    Sample: CRC

Fig. 1| SCD1 is upregulated in human CRC tissues and associated with CRC prognosis. a SCD1 mRNA level in colorectal cancer tissues (CRC) and matched adjacent non-tumor tissues (Control) detected by Real Time-PCR. b Representative Western blot and quantification data of SCD1 and EMT markers (E-cadherin and vimentin) in colorectal cancer tissue and matched adjacent non-tumor tissue.

4). Extracellular vesicles derived from mesenchymal stem cells alleviate neuroinflammation and mechanical allodynia in interstitial cystitis rats by inhibiting NLRP3 inflammasome activation. Journal of Neuroinflammation, 2022 (PubMed: 35387668) [IF=9.3]

Application: WB    Species: rat    Sample: spinal dorsal horn

Fig. 8 | MSC-EVs inhibit activity of TLR4/NF-κB signal pathway in SDH of IC rats. A–C Western blot analysis showing that expression level of TLR4 and phosphorylation ratio of NF‐κB (p65) were signifcantly increased in SDH of IC rats compared with normal rats, and intrathecal injection of MSC-EVs signifcantly decreased expression level of TLR4 and phosphorylation ratio of NF‐κB (p65) in SDH of IC rats

5). FOSL1 promotes tumorigenesis in colorectal carcinoma by mediating the FBXL2/Wnt/β-catenin axis via Smurf1. Pharmacological Research, 2021 (PubMed: 33450386) [IF=9.3]

Application: WB    Species: human    Sample: RKO and SW480 cells

Fig. 6| .Smurf1 silencing inhibits malignant biological behavior and angiogenesis in CC cells. RKO and SW480 cells were co-transfected with shFosl1 +Lv-Smurf1 or Lv-Fosl1 +shSmurf1. A, mRNA expression of Smurf1 in cells determined by RT-qPCR; B, protein expression of Smurf1 in cells evaluated by western blot

6). EGFR tyrosine kinase activity and Rab GTPases coordinate EGFR trafficking to regulate macrophage activation in sepsis. Cell Death & Disease, 2022 (PubMed: 36344490) [IF=9.0]

7). The mmu_circRNA_37492/hsa_circ_0012138 function as potential ceRNA to attenuate obstructive renal fibrosis. Cell Death & Disease, 2022 (PubMed: 35246505) [IF=9.0]

Application: WB    Species: Mouse    Sample: BUMPT cells

Fig. 2: Opposite fibrotic effects in circRNA_37492 inhibition & overexpression. BUMPT cells were pre-transfected with either scrambled siRNA or siRNA circRNA_37492, scramble plasmids, or circRNA_37492 plasmids and co-treated with or without TGF-β1 for 24 h. A Type I, III collagen and FN expression by western blot analysis. B Quantification of protein band of type I collagen, type III collagen, and FN by grayscale analysis. C Type I, III collagen and FN expression by western blot analysis. D Quantification of protein band of type I collagen, type III collagen, and FN by grayscale analysis. Quantitative data are presented as the mean ± SD (n = 6 per group). *p 

8). Overexpression of miR-92a attenuates kidney ischemia–reperfusion injury and improves kidney preservation by inhibiting MEK4/JNK1-related autophagy. Cellular & Molecular Biology Letters, 2023 (PubMed: 36890442) [IF=8.3]

Application: WB    Species: Mouse    Sample:

Fig. 1 IC and IR downregulated the expression of miR-92a and increased apoptosis and autophagy in mice. A The content of serum Cr and BUN in the sham and IR48 groups. BThe expression of miR-92a. Representative band C and statistical analysis D of LC3B II/I and active caspase 3 expression using Western blotting. E Representative images of H&E staining (×100), TUNEL (×100), and TEM (×2000) in each group. F The statistical results of autophagosomes in I, IC, and IR groups. Using Western blotting, the representative band G and statistical analysis H of LC3B II/I, Beclin 1, pJNK, JNK, MEK4, and active caspase 3 expression. *P 

9). Amelioration of nonalcoholic fatty liver disease by swertiamarin in fructose-fed mice. Phytomedicine, 2019 (PubMed: 31005808) [IF=7.9]

10). SERPINA1 is a hub gene associated with intervertebral disc degeneration grade and affects the nucleus pulposus cell phenotype through the ADIRF-AS1/miR-214-3p axis. Translational Research, 2022 (PubMed: 35196590) [IF=7.8]

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