Recombinant
RabMAb

Anti-GAPDH antibody [EPR16891] (ab181602)

Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab181602 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/500.
Flow Cyt 1/180.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/60.

Target

  • FunctionHas 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 (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.
  • PathwayCarbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 1/5.
  • Sequence similaritiesBelongs to the glyceraldehyde-3-phosphate dehydrogenase family.
  • Post-translational
    modifications
    S-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus.
    ISGylated.
  • Cellular localizationCytoplasm > cytosol. Nucleus. Cytoplasm > perinuclear region. Membrane. Translocates to the nucleus following S-nitrosylation and interaction with SIAH1, which contains a nuclear localization signal (By similarity). Postnuclear and Perinuclear regions.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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
    see all

Anti-GAPDH antibody [EPR16891] images

  • ab181602 staining GAPDH in undifferentiated human embryonic stem cells on a murine embryonic fibroblast (MEFs) feeding layer by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized and blocked with 1% serum for 30 minutes at room temperature. Samples were incubated with primary antibody (1/100 in 1% serum, 0.1% triton, 0.1% BSA + PBS) for 16 hours at 4°C. An Alexa Flour® 488-conjugated donkey anti-rabbit IgG polyclonal was used as the secondary antibody at a dilution of 1/500.

    See Abreview

  • All lanes : Anti-GAPDH antibody [EPR16891] (ab181602) at 1/10000 dilution

    Lane 1 : Mouse kidney lysates
    Lane 2 : Mouse spleen lysates
    Lane 3 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysates
    Lane 4 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
    Lane 5 : Rat brain lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size : 36 kDa
    Observed band size : 36 kDa

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling GAPDH with ab181602 at 1/2000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus and cytoplasmic staining on lymphocyte of rat spleen is observed. Counter stained with Hematoxylin.

     

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  • Immunocytochemistry/immunofluorescence staining of 4% paraformaldehyde fixed; 0.1% triton X 100 permeabilized HeLa (human cervix adenocarcinoma) cells labelling GAPDH with ab181602 at dilution of 1/500. The secondary antibody used was Alexa Fluor® 488; goat anti-rabbit IgG (ab150077) at a dilution of 1/400. Nucleus was counter-stained with DAPI (blue). ab7291, a mouse anti-tubulin antibody (1/500) was used to stain tubulin along with ab150120 (AlexaFluor®594 goat anti-mouse secondary, 1/500). The negative controls are shown in the bottom middle and right hand panels- for negative control 1 primary antibody (ab181602; 1/500) and secondary antibody (ab150120; 1/500) was used. For negative control 2 primary antibody (ab7291; 1/500) and secondary antibody (ab150077; 1/400) was used.

  • Flow cytometric analysis of 2% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling GAPDH with ab181602 at 1/180 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

  • All lanes : Anti-GAPDH antibody [EPR16891] (ab181602) at 1/50000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
    Lane 2 : Xenopus(X. tropicalis) muscle lysates
    Lane 3 : UMNSAH/DF-1 (Transformed chicken embyronic fibroblast cells) whole cell lysates
    Lane 4 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size : 36 kDa
    Observed band size : 36 kDa

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded human transitional cell carcinoma of bladder tissue labeling GAPDH with ab181602 at 1/2000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasmic and nucleus staining on the tumor cells of transitional cell carcinoma of Human bladder is observed. Counter stained with Hematoxylin.

     

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  • All lanes : Anti-GAPDH antibody [EPR16891] (ab181602) at 1/10000 dilution

    Lane 1 : COS-1 (African green monkey kidney fibroblast-like cell line) whole cell lysates
    Lane 2 : Zebrafish lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size : 36 kDa
    Observed band size : 36 kDa

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-GAPDH antibody [EPR16891] (ab181602) at 1/10000 dilution

    Lane 1 : Human fetal brain lysates
    Lane 2 : Human fetal heart lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size : 36 kDa
    Observed band size : 36 kDa

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling GAPDH with ab181602 at 1/2000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus and cytoplasmic staining on lymphocytes of mouse spleen is observed. Counter stained with Hematoxylin.

     

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  • GAPDH was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell extract with ab181602 at 1/60 dilution. Western blot was performed from the immunoprecipitate using ab181602 at 1/5000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Lane 1: HeLa whole cell extract. Lane 2: PBS instead of HeLa whole cell extract.


    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

References for Anti-GAPDH antibody [EPR16891] (ab181602)

This product has been referenced in:
  • Gerarduzzi C  et al. Silencing SMOC2 ameliorates kidney fibrosis by inhibiting fibroblast to myofibroblast transformation. JCI Insight 2:N/A (2017). WB ; Mouse . Read more (PubMed: 28422762) »
  • Zhu H  et al. IDH1 R132H Mutation Enhances Cell Migration by Activating AKT-mTOR Signaling Pathway, but Sensitizes Cells to 5-FU Treatment as NADPH and GSH Are Reduced. PLoS One 12:e0169038 (2017). WB ; Human . Read more (PubMed: 28052098) »

See all 83 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Fibroblasts)
Gel Running Conditions Reduced Denaturing
Loading amount 50 µg
Specification Fibroblasts
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Abcam user community

Verified customer

Submitted Nov 29 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Brain, liver, retina)
Gel Running Conditions Reduced Denaturing (12% gel)
Loading amount 20 µg
Specification Brain, liver, retina
Blocking step Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
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Abcam user community

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Submitted Oct 25 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Zebrafish Tissue lysate - whole (Brain, retina)
Gel Running Conditions Reduced Denaturing (12% gel)
Loading amount 20 µg
Specification Brain, retina
Blocking step Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

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Submitted Oct 25 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (ARPE19, hfRPE, HEK293)
Gel Running Conditions Reduced Denaturing (12% gel)
Loading amount 20 µg
Specification ARPE19, hfRPE, HEK293
Blocking step Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

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Submitted Oct 25 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (skin)
Gel Running Conditions Reduced Denaturing
Loading amount 20 µg
Specification skin
Blocking step Commercial blocking buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 23°C
Username

Maral Tajerian

Verified customer

Submitted Jul 15 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (undifferentiated Human ES cells on MEFs)
Permeabilization Yes - see blocking buffer
Specification undifferentiated Human ES cells on MEFs
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: rt°C
Fixative Paraformaldehyde
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Submitted Feb 22 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HUES7 ES cells)
Gel Running Conditions Reduced Denaturing (12%)
Loading amount 20 µg
Specification HUES7 ES cells
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C
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Submitted Feb 04 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HEPG2)
Specification HEPG2
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: rt°C
Fixative Paraformaldehyde
Username

Abcam user community

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Submitted Nov 18 2015

Application Western blot
Sample Human Cell lysate - whole cell (HEPG2)
Gel Running Conditions Reduced Denaturing (12.5)
Loading amount 20 µg
Specification HEPG2
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C
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Submitted Nov 13 2015

Application Western blot
Sample Mouse Tissue lysate - nuclear (liver)
Gel Running Conditions Reduced Denaturing (gel 12%)
Loading amount 36 µg
Specification liver
Blocking step super blocker as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C
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Mr. Mathew Joseph

Verified customer

Submitted Sep 23 2015

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"