• Western blot analysis of extracts of various cell lines, using GAPDH antibody (STJ11103691) at 1:10000 dilution. Secondary antibody: HRP Goat Anti-rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% non-fat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 1s.
  • Immunofluorescence analysis of U-2 OS cells using GAPDH Rabbit polyclonal antibody (STJ11103691) at dilution of 1:100 (40x lens). Secondary antibody: Cy3 Goat Anti-Rabbit IgG (H+L) at 1:500 dilution. Blue: DAPI for nuclear staining.
  • Immunohistochemistry of paraffin-embedded rat kidney using GAPDH rabbit polyclonal antibody (STJ11103691) at dilution of 1:100 (40x lens). Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6. 0 before commencing with immunohistochemistry staining protocol.
  • Immunofluorescence analysis of NIH-3T3 cells using GAPDH Rabbit polyclonal antibody (STJ11103691) at dilution of 1:100 (40x lens). Secondary antibody: Cy3 Goat Anti-Rabbit IgG (H+L) at 1:500 dilution. Blue: DAPI for nuclear staining.
  • Immunohistochemistry of paraffin-embedded rat pancreas using GAPDH rabbit polyclonal antibody (STJ11103691) at dilution of 1:100 (40x lens). Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6. 0 before commencing with immunohistochemistry staining protocol.
  • Immunofluorescence analysis of C6 cells using GAPDH Rabbit polyclonal antibody (STJ11103691) at dilution of 1:100 (40x lens). Secondary antibody: Cy3 Goat Anti-Rabbit IgG (H+L) at 1:500 dilution. Blue: DAPI for nuclear staining.
  • Immunohistochemistry of paraffin-embedded human liver cancer using GAPDH rabbit polyclonal antibody (STJ11103691) at dilution of 1:100 (40x lens). Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6. 0 before commencing with immunohistochemistry staining protocol.
  • Immunohistochemistry analysis of GAPDH in paraffin-embedded mouse kidney using GAPDH Rabbit polyclonal antibody (STJ11103691) at dilution of 1:100 (40x lens). Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6. 0 before commencing with immunohistochemistry staining protocol.
  • Immunohistochemistry of paraffin-embedded mouse lung using GAPDH rabbit polyclonal antibody (STJ11103691) at dilution of 1:100 (40x lens). Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6. 0 before commencing with immunohistochemistry staining protocol.
  • Immunohistochemistry analysis of GAPDH in paraffin-embedded human liver cancer using GAPDH Rabbit polyclonal antibody (STJ11103691) at dilution of 1:100 (40x lens). Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6. 0 before commencing with immunohistochemistry staining protocol.
  • Immunofluorescence analysis of C6 cells using GAPDH rabbit polyclonal antibody (STJ11103691) at dilution of 1:100 (40x lens). Blue: DAPI for nuclear staining.
  • Immunohistochemistry analysis of GAPDH in paraffin-embedded rat kidney using GAPDH Rabbit polyclonal antibody (STJ11103691) at dilution of 1:100 (40x lens). Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6. 0 before commencing with immunohistochemistry staining protocol.
  • Immunofluorescence analysis of NIH-3T3 cells using GAPDH rabbit polyclonal antibody (STJ11103691) at dilution of 1:100 (40x lens). Blue: DAPI for nuclear staining.
  • Western blot analysis of various lysates using GAPDH Rabbit polyclonal antibody (STJ11103691) at 1:10000 dilution. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (STJS000856) at 1:10000 dilution. Lysates/proteins: 25 Mu g per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 1s.
  • Immunofluorescence analysis of U-2 OS cells using GAPDH rabbit polyclonal antibody (STJ11103691) at dilution of 1:100 (40x lens). Blue: DAPI for nuclear staining.

Anti-GAPDH antibody (1-335) (STJ11103691)

SKU:
STJ11103691

Current Stock:
Host: Rabbit
Applications: WB/IHC/IF/IP
Reactivity: Human/Mouse/Rat
Note: STRICTLY FOR FURTHER SCIENTIFIC RESEARCH USE ONLY (RUO). MUST NOT TO BE USED IN DIAGNOSTIC OR THERAPEUTIC APPLICATIONS.
Short Description: Rabbit polyclonal antibody anti-GAPDH (1-335) is suitable for use in Western Blot, Immunohistochemistry, Immunofluorescence and Immunoprecipitation research applications.
Clonality: Polyclonal
Conjugation: Unconjugated
Isotype: IgG
Formulation: PBS with 0.01% Thimerosal, 50% Glycerol, pH7.3.
Purification: Affinity purification
Dilution Range: WB 1:2000-1:10000
IHC-P 1:50-1:200
IF/ICC 1:50-1:200
IP 1:50-1:100
Storage Instruction: Store at-20°C for up to 1 year from the date of receipt, and avoid repeat freeze-thaw cycles.
Gene Symbol: GAPDH
Gene ID: 2597
Uniprot ID: G3P_HUMAN
Immunogen Region: 1-335
Immunogen: Recombinant fusion protein containing a sequence corresponding to amino acids 1-335 of human GAPDH (NP_002037.2).
Immunogen Sequence: MGKVKVGVNGFGRIGRLVTR AAFNSGKVDIVAINDPFIDL NYMVYMFQYDSTHGKFHGTV KAENGKLVINGNPITIFQER DPSKIKWGDAGAEYVVESTG VFTTMEKAGAHLQGGAKRVI ISAPSADAPMFVMGVNHEKY DNSLKIISNASCTTNCLAPL AKVIHDNFGIVEGLMTTVHA ITATQKTVDGPSGKLWRDGR GALQNIIPASTGAAKAVGKV IPELNGKLTGMAFRVPTAN
Post Translational Modifications S-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus. 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. (Microbial infection) Glycosylated by C.rodentium protein NleB, enteropathogenic E.coli protein NleB1 and S.typhimurium protein Ssek1: arginine GlcNAcylation prevents the interaction with TRAF2 and TRAF3. This leads to reduced ubiquitination of TRAF2 and TRAF3, and subsequent inhibition of NF-kappa-B signaling and type I interferon production, respectively.
Function Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. 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. 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. 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. Also plays a role in innate immunity by promoting TNF-induced NF-kappa-B activation and type I interferon production, via interaction with TRAF2 and TRAF3, 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.
Protein Name Glyceraldehyde-3-Phosphate Dehydrogenase
Gapdh
Peptidyl-Cysteine S-Nitrosylase Gapdh
Database Links Reactome: R-HSA-70171
Reactome: R-HSA-70263
Cellular Localisation Cytoplasm
Cytosol
Nucleus
Perinuclear Region
Membrane
Cytoskeleton
Translocates To The Nucleus Following S-Nitrosylation And Interaction With Siah1
Which Contains A Nuclear Localization Signal
Postnuclear And Perinuclear Regions
Alternative Antibody Names Anti-Glyceraldehyde-3-Phosphate Dehydrogenase antibody
Anti-Gapdh antibody
Anti-Peptidyl-Cysteine S-Nitrosylase Gapdh antibody
Anti-GAPDH antibody
Anti-GAPD antibody
Anti-CDABP0047 antibody
Anti-OK antibody
Anti-SW-cl.12 antibody

Information sourced from Uniprot.org

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