ARG52320

anti-GAPDH antibody [1D4]

anti-GAPDH antibody [1D4] for ICC/IF,Western blot and Human,Monkey,Mouse,Rat

Cancer antibody; Controls and Markers antibody; Metabolism antibody; Neuroscience antibody; Signaling Transduction antibody; Loading Control antibody; Loading Control antibody for Cytoplasmic Fractions; Organelle Marker antibody for Cytoplasm; Autophagy Study antibody

Overview

Product Description Mouse Monoclonal antibody [1D4] recognizes GAPDH
Tested Reactivity Hu, Ms, Rat, Mk
Predict Reactivity Bird, Bov
Tested Application ICC/IF, WB
Host Mouse
Clonality Monoclonal
Clone 1D4
Isotype IgM
Target Name GAPDH
Antigen Species Pig
Immunogen Purified porcine 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/IF1:100
WB1:1000
Application Note Specific for the ~38k Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) protein.
* The dilutions indicate recommended starting dilutions and the optimal dilutions or concentrations should be determined by the scientist.

Properties

Form Liquid
Purification Total IgM fraction--concentrated culture supernatant
Buffer Total IgM fraction and 10 mM Sodium azide
Preservative 10 mM Sodium azide
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 or below. 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 GAPDH 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 GAPDH 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. 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]
Research Area Cancer antibody; Controls and Markers antibody; Metabolism antibody; Neuroscience antibody; Signaling Transduction antibody; Loading Control antibody; Loading Control antibody for Cytoplasmic Fractions; Organelle Marker antibody for Cytoplasm; Autophagy Study antibody
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.

Images (4) Click the Picture to Zoom In

  • ARG52320 anti-GAPDH antibody [1D4] ICC/IF image

    Immunofluorescence: HeLa cells stained with ARG52320 anti-GAPDH antibody [1D4] (green) at 1:100 dilution. DAPI (blue) for nuclear staining.

  • ARG52320 anti-GAPDH antibody [1D4] WB image

    Western Blot: rat hippocampal lysate showing specific immunolabeling of the ~38k GAPDH protein stained with anti-GAPDH antibody [1D4] (ARG52320).

  • ARG52320 anti-GAPDH antibody [1D4] ICC/IF image

    Immunofluorescence: Human neuroblastoma SH-SY5Y cells stained withARG52320 anti-GAPDH (green) and ARG52347 anti-Neurofilament NF-H antibody (red) and DNA (blue).

  • ARG52320 anti-GAPDH antibody [1D4] WB image

    Western blot: HEK293, HeLa, SH-SY5Y, COS1, NIH/3T3 and C6 cell lysates stained with ARG52320 anti-GAPDH antibody [1D4] at 1:2000 dilution.

Clone References

A functional rhodopsin-green fluorescent protein fusion protein localizes correctly in transgenic Xenopus laevis retinal rods and is expressed in a time-dependent pattern.

WB / Bovine

Moritz OL et al.
J Biol Chem.,  (2001)

publication_link

 

hr_line

Disease sequence from mutant rhodopsin allele to rod and cone photoreceptor degeneration in man.

WB / Human

Cideciyan AV et al.
Proc Natl Acad Sci U S A.,  (1998)

publication_link

 

hr_line

Monoclonal antibodies to rhodopsin: characterization, cross-reactivity, and application as structural probes.

WB / Amphibian, Bovine

Molday RS et al.
Biochemistry.,  (1983)

publication_link

 

hr_line

Antigen-antibody interaction. Synthetic peptides define linear antigenic determinants recognized by monoclonal antibodies directed to the cytoplasmic carboxyl terminus of rhodopsin.

IP / Bovine

Hodges RS et al.
J Biol Chem.,  (1988)

publication_link

 

hr_line