PRODUCT CODE: EM1101

GAPDH Mouse Monoclonal Antibody [5-E10] (EM1101)

  • Zebrafish

Applications

  • WB

  • ICC

REACTIVITY

  • Human

  • Rat

  • Zebrafish

  • Rabbit

Western blot analysis of GAPDH on different cell lysates using anti-GAPDH antibody at 1/5000 dilution.<br />
Positive control:<br />
Lane 1: HepG2<br />
Lane 2: Hela<br />
Lane 3: PC12<br />
Lane 4: NIH/3T3<br />
Lane 5: MCF-7<br />
Lane 6: Rabbit liver<br />
Lane 7: Zebrafish
  • Western blot analysis of GAPDH on different cell lysates using anti-GAPDH antibody at 1/5000 dilution.<br />
Positive control:<br />
Lane 1: HepG2<br />
Lane 2: Hela<br />
Lane 3: PC12<br />
Lane 4: NIH/3T3<br />
Lane 5: MCF-7<br />
Lane 6: Rabbit liver<br />
Lane 7: Zebrafish
  • ICC staining GAPDH in F9 cells (red). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
  • ICC staining GAPDH in Hela cells (red). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
  • ICC staining GAPDH in HepG2 cells (red). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
Western blot analysis of GAPDH on different cell lysates using anti-GAPDH antibody at 1/5000 dilution.
Positive control:
Lane 1: HepG2
Lane 2: Hela
Lane 3: PC12
Lane 4: NIH/3T3
Lane 5: MCF-7
Lane 6: Rabbit liver
Lane 7: Zebrafish

Applications

  • WB

  • ICC

REACTIVITY

  • Human

  • Rat

  • Zebrafish

  • Rabbit

SPECIFICATIONS

Product Type

Mouse monoclonal primary

Product Name

GAPDH Mouse Monoclonal Antibody [5-E10] (EM1101)

Immunogen

Synthetic peptide within human gapdh aa 10-50.

Host

Mouse

Positive Control

HepG2, Hela, PC12, NIH/3T3, MCF-7, Rabbit liver, zebrafish, F9

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

5-E10

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, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

2 ug/ul

PURIFICATION

Protein L purified.

MOLECULAR WEIGHT

36 kDa

Isotype

IgM

APPLICATION DILUTION

  • WB

  • 1:5,000-1:10,000

  • ICC

  • 1:200-1:500

TARGET

UNIPROT #

PROTEIN NAME

GAPDH

SYNONYMS

38 kDa BFA-dependent ADP-ribosylation substrate antibody; aging associated gene 9 protein antibody; Aging-associated gene 9 protein antibody; BARS-38 antibody; cb609 antibody; EC 1.2.1.12 antibody; Epididymis secretory sperm binding protein Li 162eP antibody; G3P_HUMAN antibody; G3PD antibody; G3PDH antibody; GAPD antibody; GAPDH antibody; Glyceraldehyde 3 phosphate dehydrogenase antibody; Glyceraldehyde-3-phosphate dehydrogenase antibody; HEL-S-162eP antibody; KNC-NDS6 antibody; MGC102544 antibody; MGC102546 antibody; MGC103190 antibody; MGC103191 antibody; MGC105239 antibody; MGC127711 antibody; MGC88685 antibody; OCAS, p38 component antibody; OCT1 coactivator in S phase, 38-KD component antibody; peptidyl cysteine S nitrosylase GAPDH antibody; Peptidyl-cysteine S-nitrosylase GAPDH antibody; wu:fb33a10 antibody

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

FUNCTION

GAPDH (Glyceraldehyde-3-phosphate dehydrogenase) has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. It participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis. GAPDH 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.

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