PRODUCT CODE: R1210-1

GAPDH Rabbit Polyclonal Antibody (R1210-1)

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

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Western blot analysis of GAPDH on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used at a 1:5,000 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: F9 cell lysate, untreated <br />
Lane 2: NCCIT cell lysate, untreated<br />
Lane 3: PC-12 cell lysate, untreated
  • Western blot analysis of GAPDH on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used at a 1:5,000 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: F9 cell lysate, untreated <br />
Lane 2: NCCIT cell lysate, untreated<br />
Lane 3: PC-12 cell lysate, untreated
  • ICC staining GAPDH in A549 cells (green). 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 antibody (R1210-1) at a dilution of 1:100 for 1 hour at room temperature, washed with PBS. Alexa Fluorc™ 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining GAPDH in LOVO cells (green). 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 antibody (R1210-1) at a dilution of 1:100 for 1 hour at room temperature, washed with PBS. Alexa Fluorc™ 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining GAPDH in MCF-7 cells (green). 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 antibody (R1210-1) at a dilution of 1:100 for 1 hour at room temperature, washed with PBS. Alexa Fluorc™ 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded rat kidney tissue using anti-GAPDH antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (R1210-1) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue using anti-GAPDH antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (R1210-1) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-GAPDH antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (R1210-1) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded mouse testis tissue using anti-GAPDH antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (R1210-1) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of GAPDH was done on Hela cells. The cells were fixed, permeabilized and stained with GAPDH antibody at 1/100 dilution (blue) compared with an unlabelled control (cells without incubation with primary antibody; red). After incubation of the primary antibody on room temperature for an hour, the cells was stained with a Alexa Fluor™ 488-conjugated goat anti-rabbit IgG Secondary antibody at 1/500 dilution.
Western blot analysis of GAPDH on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used at a 1:5,000 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: F9 cell lysate, untreated
Lane 2: NCCIT cell lysate, untreated
Lane 3: PC-12 cell lysate, untreated

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Rabbit polyclonal primary

Product Name

GAPDH Rabbit Polyclonal Antibody (R1210-1)

Immunogen

This antibody is produced by immunizing rabbits with full length recombinant protein of gapdh.

Host

Rabbit

Positive Control

F9, NCCIT, PC-12, A549, LOVO, MCF-7, rat kidney tissue, human colon cancer tissue, human spleen tissue, mouse testis tissue, Hela.

Conjugation

Unconjugated

Clonality

Polyclonal

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C. 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 affinity purified.

MOLECULAR WEIGHT

36 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB:1:5,000-1:10,000

  • ICC:1:100-1:200

  • IHC-P:1:50-1:200

  • FC:1:50-1:100

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

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) catalyzes the phosphorylation of glyceraldehyde-3-phosphate during glycolysis. It participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis. GAPDH is thought to be a constitutively expressed housekeeping protein. For this reason, GAPDH mRNA and protein levels are often measured as controls in experiments quantifying specific changes in expression of other targets.

CITATIONS

  • Zan, Jie et al.

    Rabies virus matrix protein induces apoptosis by targeting mitochondria. | Experimental Cell Research [2016]

  • Zan, Jie et al.

    Rabies Virus Infection Induces Microtubule Depolymerization to Facilitate Viral RNA Synthesis by Upregulating HDAC6. | Frontiers in Cellular and Infection Microbiology [2017]

  • Wang, Ke et al.

    Combination of CALR and PDIA3 is a potential prognostic biomarker for non-small cell lung cancer. | Oncotarget [2017]

  • Lei, Zhong et al.

    PARK2 inhibits osteosarcoma cell growth through the JAK2/STAT3/VEGF signaling pathway. | Cell Death & Disease [2018]

  • Feng, Chen-Zhuo et al.

    Dendrobium polysaccharides attenuate cognitive impairment in senescence-accelerated mouse prone 8 mice via modulation of microglial activation. | Brain Research [2019]