Overview

  • Product nameAnti-GAPDH [6C5] antibodySee all GAPDH primary antibodies ...
  • Description
    Mouse monoclonal [6C5] to GAPDH
  • SpecificityThis antibody can be used as a loading control antibody, however we also recommend ab9484 as an alternative. GAPDH is a 146 kDa tetramer composed of four 30-40 kDa subunits. There is no cross-reaction with GAPDH from yeast. Preliminary data indicates that the antibody recognizes the monomer (36 kDa) and also the dimer forms of GAPDH, but not the tetrameric form of the protein.
  • Tested applicationsELISA, ICC, WB, ICC/IF, IHC-Fr more details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Chicken, Hamster, Cat, Dog, Human, Pig, Xenopus laevis, Fruit fly (Drosophila melanogaster), Fish, Monkey, Zebrafish, Baboon, African Green Monkey
    Predicted to work with: Horse, Guinea pig, Xenopus tropicalis

    Does not react with

    Goat, Cow, Saccharomyces cerevisiae
  • Immunogen

    Rabbit muscle GAPDH.

Properties

Applications

Our Abpromise guarantee covers the use of ab8245 in the following tested applications.

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

Application Notes
ELISA Use at an assay dependent dilution.
ICC Use at an assay dependent dilution.
WB 1/500 - 1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 40.2 kDa). PubMed: 16450009
ICC/IF Use at an assay dependent dilution.
IHC-Fr 1/500.

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.
  • Target information above from: UniProt accession P04406 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    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
    • G3P_HUMAN antibody
    • G3PD antibody
    • G3PDH antibody
    • GAPD antibody
    • GAPDH antibody
    • GAPDH antibody
    • Glyceraldehyde 3 phosphate dehydrogenase antibody
    • Glyceraldehyde 3 phosphate dehydrogenase liver antibody
    • Glyceraldehyde 3 phosphate dehydrogenase muscle antibody
    • Glyceraldehyde-3-phosphate dehydrogenase 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 [6C5] antibody images

  • All lanes : Anti-GAPDH [6C5] antibody (ab8245) at 20000

    Lane 1 : Mouse hippocampus whole cell lysate
    Lane 2 : Rat hippocampus whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP-conjugated Rabbit anti-mouse at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 40.2 kDa
    Observed band size : 36 kDa (why is the actual band size different from the predicted?)


    Exposure time : 10 seconds

    This image is courtesy of an anonymous Abreview

    See Abreview

  • All lanes : Anti-GAPDH [6C5] antibody (ab8245) at 2.5 µg/ml

    Lane 1 : HeLa Nuclear
    Lane 2 : HeLa whole cell lysate
    Lane 3 : A431 cell lysate
    Lane 4 : Jurkat cell lysate
    Lane 5 : HEK293 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Alexa Fluor anti-mouse at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size : 40.2 kDa
    Observed band size : 37 kDa (why is the actual band size different from the predicted?)
    Fluorescence detection of secondary antibody.
  • All lanes : Anti-GAPDH [6C5] antibody (ab8245) at 1/20000 dilution

    Lane 1 : Rabbit aorta, whole tissue lysate at 10 µg
    Lane 2 : Blank
    Lane 3 : Mouse aorta, whole tissue lysate at 5 µg
    Lane 4 : Mouse aorta, whole tissue lysate at 7.5 µg
    Lane 5 : Mouse aorta, whole tissue lysate at 10 µg
    Lane 6 : Mouse aorta, whole tissue lysate at 12.5 µg
    Lane 7 : Mouse aorta, whole tissue lysate at 15 µg

    Secondary
    HRP conjugated sheep anti-mouse antibody at 1/10000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 40.2 kDa


    Exposure time : 30 seconds

    This image is courtesy of an anonymous Abreview

    See Abreview



  • Predicted band size : 40.2 kDa

    20 µg of Raji whole cell lysate (ab7908) probed with mouse monoclonal to GAPDH (ab8245) at 10µg/ml.

    20 µg of Raji whole cell lysate (ab7908) probed with mouse monoclonal to GAPDH (ab8245) at 10µg/ml.

References for Anti-GAPDH [6C5] antibody (ab8245)

This product has been referenced in:
  • Toret CP  et al. A genome-wide screen identifies conserved protein hubs required for cadherin-mediated cell-cell adhesion. J Cell Biol 204:265-79 (2014). WB ; Fruit fly (Drosophila melanogaster) . Read more (PubMed: 24446484) »
  • Zhao L  et al. Notch signaling regulates cardiomyocyte proliferation during zebrafish heart regeneration. Proc Natl Acad Sci U S A 111:1403-8 (2014). Zebrafish . Read more (PubMed: 24474765) »

See all 216 Publications for this product

Product Wall

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing (4-12%)
Sample Human Cell lysate - whole cell (Colon cancer)
Specification Colon cancer
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Submitted Apr 14 2014

Application ELISA
Sample Human Recombinant protein (HEK293)
Specification HEK293
Type Direct
Blocking step Superblock in PBS (Pierce Cat#37580) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10µg/mL · Temperature: 20°C
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Submitted Mar 17 2014

Application Western blot
Loading amount 10000 cells
Gel Running Conditions Reduced Denaturing (6%)
Sample Spodoptera frugiperda (SF9 cells) Cell lysate - whole cell (293 cells and Sf9 cells)
Specification 293 cells and Sf9 cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
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Submitted Feb 18 2014

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (15% Gel)
Sample Human Tissue lysate - whole (Vastus lateralis muscle)
Specification Vastus lateralis muscle
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Feb 18 2014

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (12% gel)
Sample Mouse Tissue lysate - whole (Skeletal muscle)
Specification Skeletal muscle
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
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Submitted Jan 09 2014

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (4-12%)
Sample Human Cell lysate - whole cell (HCT116)
Specification HCT116
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Submitted Dec 20 2013

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (10)
Sample Dog Cell lysate - whole cell (MDCK)
Specification MDCK
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Oct 28 2013

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (10)
Sample Syrian Hamster Cell lysate - whole cell (BHK)
Specification BHK
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Oct 28 2013

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (10)
Sample African Green Monkey Cell lysate - whole cell (COS)
Specification COS
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Oct 24 2013

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (HEK293)
Specification HEK293
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
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Submitted Oct 18 2013

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