PRODUCT CODE: ER1706-83

GAPDH Antibody (ER1706-83)

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of GAPDH on different lysates using anti-GAPDH antibody at 1/2,000 dilution.<br />
 Positive control:<br />
 Lane 1: PC-12 <br />
 Lane 2: L929<br />
 Lane 3: NIH/3T3 <br />
 Lane 4: F9<br />
 Lane 5: Mouse liver tissue<br />
  • Western blot analysis of GAPDH on different lysates using anti-GAPDH antibody at 1/2,000 dilution.<br />
 Positive control:<br />
 Lane 1: PC-12 <br />
 Lane 2: L929<br />
 Lane 3: NIH/3T3 <br />
 Lane 4: F9<br />
 Lane 5: Mouse liver tissue<br />
  • ICC staining GAPDH in LOVO cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
  • Immunohistochemical analysis of paraffin-embedded human pancreas tissue using anti-GAPDH antibody. Counter stained with hematoxylin.
  • Immunohistochemical analysis of paraffin-embedded mouse testis tissue using anti-GAPDH antibody. Counter stained with hematoxylin.
  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-GAPDH antibody. Counter stained with hematoxylin.
  • Flow cytometric analysis of MCF-7 cells with GAPDH antibody at 1/50 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody.
Western blot analysis of GAPDH on different lysates using anti-GAPDH antibody at 1/2,000 dilution.
Positive control:
Lane 1: PC-12
Lane 2: L929
Lane 3: NIH/3T3
Lane 4: F9
Lane 5: Mouse liver tissue

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Rabbit polyclonal primary

Product Name

GAPDH Antibody (ER1706-83)

Immunogen

Peptide

Host

Rabbit

Positive Control

PC-12, L929, NIH/3T3, F9, mouse liver tissue lysate, human tonsil tissue, human pancreas tissue, mouse testis tissue, MCF-7.

Conjugation

Unconjugated

Clonality

Polyclonal

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Peptide affinity purified

MOLECULAR WEIGHT

36 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:2,000-5,000

  • ICC

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1

  • 100

TARGET

UNIPROT #

PROTEIN NAME

Glyceraldehyde-3-phosphate dehydrogenase

GENE NAME

GAPDH

SYNONYMS

GAPDH, GAPD, CDABP0047, OK/SW-cl.12

SEQUENCE SIMILARITIES

Belongs to the glyceraldehyde-3-phosphate dehydrogenase family.

POST-TRANSLATIONAL MODIFICATION

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. 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.

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.