PRODUCT CODE: M1310-2

GAPDH Mouse Monoclonal Antibody [12D6] (M1310-2)

  • Zebrafish

Applications

  • WB

  • ICC

REACTIVITY

  • Human

  • Mouse

  • Rat

  • Zebrafish

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Western blot analysis of GAPDH on different cells lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (M1210-2, 1/1000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Hela cell lysate<br />
Lane 2: A549 cell lysate<br />
Lane 3: HepG2 cell lysate<br />
Lane 4: PC-12 cell lysate<br />
Lane 5: F9 cell lysate
  • Western blot analysis of GAPDH on different cells lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (M1210-2, 1/1000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Hela cell lysate<br />
Lane 2: A549 cell lysate<br />
Lane 3: HepG2 cell lysate<br />
Lane 4: PC-12 cell lysate<br />
Lane 5: F9 cell lysate
  • ICC staining of GAPDH in D3 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (M1310-2, 1/100) for 1 hour at room temperature, washed with PBS. Alexa Fluor®555 Goat anti-Mouse IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of GAPDH in A431 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (M1310-2, 1/100) for 1 hour at room temperature, washed with PBS. Alexa Fluor®555 Goat anti-Mouse IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
  • Western blot analysis of GAPDH on zebrafish tissue lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (M1210-2, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.
Western blot analysis of GAPDH on different cells lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (M1210-2, 1/1000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Hela cell lysate
Lane 2: A549 cell lysate
Lane 3: HepG2 cell lysate
Lane 4: PC-12 cell lysate
Lane 5: F9 cell lysate

Applications

  • WB

  • ICC

REACTIVITY

  • Human

  • Mouse

  • Rat

  • Zebrafish

SPECIFICATIONS

Product Type

Mouse monoclonal primary

Product Name

GAPDH Mouse Monoclonal Antibody [12D6] (M1310-2)

Immunogen

Synthetic peptide within human gapdh aa 180-220.

Host

Mouse

Positive Control

Hela, A549, HepG2, PC-12, F9, D3, A431, zebrafish tissue lysates.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

12D6

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

1 ug/ul

PURIFICATION

Protein G purified.

MOLECULAR WEIGHT

36 kDa

Isotype

IgG1

APPLICATION DILUTION

  • WB

  • 1:2000-1:5,000

  • ICC

  • 1:200

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, nucleus.

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. 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 and suppresses their translation.

CITATIONS

  • Liu, Jiang et al.

    A critical role of DDRGK1 in endoplasmic reticulumhomoeostasis via regulation of IRE1a stability. | Nature Communications [2017]

  • Yin, Jianhua et al.

    CD44 inhibition attenuates EGFR signaling and enhances cisplatin sensitivity in human EGFR wild-type non-small-cell lung cancer cells. | International Journal of Molecular Medicine [2020]

  • Xu, Xiaodong et al.

    Upregulation of miRNA-301a-3p promotes tumor progression in gastric cancer by suppressing NKRF and activating NF-κB signaling. | International Journal of Oncology [2020]