Overview

  • Product nameAnti-GAPDH antibody [6C5]
    See 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-Frmore 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.

  • Positive control
    • ICC/IF: HeLa cells, NIH3T3 cells, SV40LT-SMC cells

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 Abreviews Notes
ELISA Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
WB 1/500 - 1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 40.2 kDa). PubMed: 16450009
ICC/IF Use a concentration of 1 - 5 µg/ml.
IHC-Fr 1/500.

Target

Anti-GAPDH antibody [6C5] images

  • All lanes : Anti-GAPDH antibody [6C5] (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.
  • ab8245 staining GAPDH in SV40LT-SMC cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab8245 at 5μg/ml and ab202272 at 1/250 overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-Mouse AlexaFluor® 488 (ab150117) (shown in green). Nuclear DNA was labelled in blue with DAPI.
    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • ab8245 staining GAPDH in NIH3T3 cells. The cells were fixed with 4% formaldehyde (10min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab8245 at 1μg/ml and ab202272 at 1/250 overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-Mouse AlexaFluor® 488 (ab150117) (shown in green). Nuclear DNA was labelled in blue with DAPI.
    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • ab8245 staining GAPDH in HeLa cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab8245 at 5μg/ml and ab6046 at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-Mouse AlexaFluor® 488 (ab150117) at 2 μg/ml (shown in green) and anti-Rabbit AlexaFluor® 594 (ab150088) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.

    Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

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

This product has been referenced in:
  • Durk MR  et al. Vitamin D receptor activation induces P-glycoprotein and increases brain efflux of quinidine: an intracerebral microdialysis study in conscious rats. Pharm Res 32:1128-40 (2015). Read more (PubMed: 25319098) »
  • Lundby A  et al. Surface-expressed insulin receptors as well as IGF-I receptors both contribute to the mitogenic effects of human insulin and its analogues. J Appl Toxicol 35:842-50 (2015). Read more (PubMed: 25413577) »

See all 348 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 (bladder)
Gel Running Conditions Reduced Denaturing (gel 10%)
Loading amount 30 µg
Treatment lentivirus for gene knock-out
Specification bladder
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Mr. Jinwoo Ahn

Verified customer

Submitted May 20 2016

Application Western blot
Sample Human Cell lysate - whole cell (Huh7 cells)
Gel Running Conditions Reduced Denaturing
Loading amount 20 µg
Specification Huh7 cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Miss. Virginia Gutiérrez

Verified customer

Submitted May 16 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Brain)
Gel Running Conditions Reduced Denaturing (4-20%)
Loading amount 12 µg
Specification Brain
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Apr 02 2016

Application Western blot
Sample Rat Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions Reduced Denaturing (12%)
Loading amount 20 µg
Specification Skeletal muscle
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Mar 17 2016

Application Western blot
Sample Human Cell lysate - whole cell (HEK293)
Gel Running Conditions Reduced Denaturing
Loading amount 15 µg
Specification HEK293
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

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Submitted Sep 09 2015

Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (rat neonatal myocytes)
Permeabilization Yes - 1% TRITON-X-100
Specification rat neonatal myocytes
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative Paraformaldehyde
Username

Abcam user community

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Submitted Jun 05 2015

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (Hela, Hep G2)
Specification Hela, Hep G2
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

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Submitted Feb 09 2015

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Cell lysate - whole cell (liver, hepatocytes)
Specification liver, hepatocytes
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

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Submitted Feb 09 2015

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Chinese Hamster Cell lysate - whole cell (CHO cell)
Specification CHO cell
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

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Submitted Oct 22 2014

Application Western blot
Loading amount 80 µg
Gel Running Conditions Reduced Denaturing (12.5% Acrylamide, 2D gel ph 3-10)
Sample Pig Tissue lysate - other (Muscle Sarcoplasm)
Specification Muscle Sarcoplasm
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Dr. Shannon Cruzen

Verified customer

Submitted Oct 06 2014

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