ARG42505

anti-GAPDH antibody

anti-GAPDH antibody for ICC/IF,IHC-Frozen sections,IHC-Formalin-fixed paraffin-embedded sections,Western blot and Human,Mouse,Rat,Dog,Monkey,Zebrafish

Overview

Product Description Goat Polyclonal antibody recognizes GAPDH
Tested Reactivity Hu, Ms, Rat, Dog, Mk, Zfsh
Tested Application ICC/IF, IHC-Fr, IHC-P, WB
Host Goat
Clonality Polyclonal
Isotype IgG
Target Name GAPDH
Antigen Species Human
Immunogen Purified recombinant peptide within aa. 240 to the C-terminus of Human GAPDH.
Conjugation Un-conjugated
Alternate Names Glyceraldehyde-3-phosphate dehydrogenase; GAPD; HEL-S-162eP; G3PD; GAPDH; Peptidyl-cysteine S-nitrosylase GAPDH; EC 2.6.99.-; EC 1.2.1.12

Application Instructions

Application Suggestion
Tested Application Dilution
ICC/IF1:50 - 1:250
IHC-Fr1:200 - 1:1000
IHC-P1:200 - 1:1000
WB1:500 - 1:5000
Application Note * The dilutions indicate recommended starting dilutions and the optimal dilutions or concentrations should be determined by the scientist.
Positive Control HT1080
Observed Size ~ 40 kDa

Properties

Form Liquid
Purification Affinity purification with immunogen.
Buffer PBS, 0.05% Sodium azide and 20% Glycerol.
Preservative 0.05% Sodium azide
Stabilizer 20% Glycerol
Concentration 2 mg/ml
Storage Instruction For continuous use, store undiluted antibody at 2-8°C for up to a week. For long-term storage, aliquot and store at -20°C. Storage in frost free freezers is not recommended. Avoid repeated freeze/thaw cycles. Suggest spin the vial prior to opening. The antibody solution should be gently mixed before use.
Note For laboratory research only, not for drug, diagnostic or other use.

Bioinformation

Database Links

GeneID: 14433 Mouse GAPDH

GeneID: 24383 Rat GAPDH

GeneID: 2597 Human GAPDH

Gene Symbol GAPDH
Gene Full Name glyceraldehyde-3-phosphate dehydrogenase
Background This gene encodes a member of the glyceraldehyde-3-phosphate dehydrogenase protein family. The encoded protein has been identified as a moonlighting protein based on its ability to perform mechanistically distinct functions. The product of this gene catalyzes an important energy-yielding step in carbohydrate metabolism, the reversible oxidative phosphorylation of glyceraldehyde-3-phosphate in the presence of inorganic phosphate and nicotinamide adenine dinucleotide (NAD). The encoded protein has additionally been identified to have uracil DNA glycosylase activity in the nucleus. Also, this protein contains a peptide that has antimicrobial activity against E. coli, P. aeruginosa, and C. albicans. Studies of a similar protein in mouse have assigned a variety of additional functions including nitrosylation of nuclear proteins, the regulation of mRNA stability, and acting as a transferrin receptor on the cell surface of macrophage. Many pseudogenes similar to this locus are present in the human genome. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Nov 2014]
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. [UniProt]
Cellular Localization 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. [UniProt]
Calculated MW 36 kDa
PTM 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. [UniProt]

Images (2) Click the Picture to Zoom In

  • ARG42505 anti-GAPDH antibody ICC/IF image

    Immunofluorescence: Hepa1-6 cells were fixed with methanol. Cells were stained with ARG42505 anti-GAPDH antibody (green) at 1:50 dilution. Nuclear staining (blue).

  • ARG42505 anti-GAPDH antibody WB image

    Western blot: 50 µg of HT1080 cell lysate stained with ARG42505 anti-GAPDH antibody at 1:2500 dilution.