Overview

  • Product name
    Anti-GAPDH antibody [6C5]
    See all GAPDH primary antibodies
  • Description
    Mouse monoclonal [6C5] to GAPDH
  • Specificity
    This 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 applications
    Suitable for: ELISA, ICC, WB, ICC/IF, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Chicken, Hamster, Cat, Dog, Human, Pig, Xenopus laevis, Fish, Monkey, Zebrafish, Baboon, African green monkey
    Predicted to work with: Horse, Guinea pig, Xenopus tropicalisDoes 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).

  • 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

  • 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 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:
  • Wang L  et al. Cannabinoid Receptor 1 Mediates Homing of Bone Marrow-Derived Mesenchymal Stem Cells Triggered by Chronic Liver Injury. J Cell Physiol 232:110-21 (2017). Read more (PubMed: 27028843) »
  • Yu N  et al. Anti-Diabetic Effects of Jiang Tang Xiao Ke Granule via PI3K/Akt Signalling Pathway in Type 2 Diabetes KKAy Mice. PLoS One 12:e0168980 (2017). Mouse . Read more (PubMed: 28045971) »

See all 811 Publications for this product

Product Wall

Application
Western blot
Sample
Rat Tissue lysate - whole (rat hippocampus tissue)
Gel Running Conditions
Non-reduced Denaturing
Loading amount
50 µg
Treatment
RIPA buffer
Specification
rat hippocampus tissue
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 Sep 14 2017

Application
Western blot
Sample
Rat Tissue lysate - whole (brain)
Gel Running Conditions
Reduced Denaturing
Loading amount
15 µg
Specification
brain
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Jun 14 2017

Application
Western blot
Sample
Bos taurus Tissue lysate - whole (Lane 1Bovine Liver and Lane 2 Bovine mammary gland)
Gel Running Conditions
Reduced Denaturing (4-15% Gradient)
Loading amount
20 µg
Specification
Lane 1Bovine Liver and Lane 2 Bovine mammary gland
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Julie Kim

Verified customer

Submitted May 12 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Mouse whole brain)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
Mouse whole 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 May 04 2017

Application
Western blot
Sample
Rat Cell lysate - whole cell (brain)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (10%)
Loading amount
15 µg
Specification
brain
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Apr 21 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (Liver)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
50 µg
Specification
Liver
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 Feb 24 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Caenorhabditis elegans Cell lysate - whole cell (whole lysates)
Gel Running Conditions
Non-reduced Denaturing
Loading amount
50 µg
Specification
whole lysates
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Joey Wu

Verified customer

Submitted Jan 25 2017

Application
Western blot
Sample
Ferret Tissue lysate - whole (Ferret whole brain tissue)
Gel Running Conditions
Reduced Denaturing (3-8% Tris-acetate gel, NuPage)
Loading amount
40 µg
Specification
Ferret whole brain tissue
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Francis Djankpa

Verified customer

Submitted Jul 11 2016

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

1-10 of 97 Abreviews or Q&A

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