製品の概要

  • 製品名
    Anti-GAPDH antibody [6C5] - Loading Control
    GAPDH 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [6C5] to GAPDH - Loading Control
  • 由来種
    Mouse
  • 特異性
    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.
  • アプリケーション
    適用あり: ELISA, ICC, WB, ICC/IF, IHC-Frmore details
  • 種交差性
    交差種: Mouse, Rat, Rabbit, Chicken, Hamster, Cat, Dog, Human, Pig, Xenopus laevis, Fish, Monkey, Zebrafish, Baboon, African green monkey
    交差が予測される動物種: Horse, Guinea pig, Xenopus tropicalis非交差種: Goat, Cow, Saccharomyces cerevisiae
  • 免疫原

    Rabbit muscle GAPDH.

  • ポジティブ・コントロール
    • ICC/IF: HeLa cells, NIH3T3 cells, SV40LT-SMC cells

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

画像

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

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) Nuclear
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 3 : A431 (Human epidermoid carcinoma cell line) cell lysate
    Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate
    Lane 5 : HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : 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 (10 minutes), 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 1 hour. 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 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) (shown in green). Nuclear DNA was labeled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

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

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : 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

    See Abreview

  • ab8245 staining GAPDH in NIH/3T3 (Mouse embryo fibroblast cell line) cells.

    The cells were fixed with 4% formaldehyde (10 minutes) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with ab8245 at 1 μg/ml and ab202272 at 1/250 overnight at +4°C, followed by a further incubation at room temperature for 1 hour with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) (shown in green). Nuclear DNA was labeled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • ab8245 staining GAPDH in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.

    The cells were fixed with 100% methanol (5 minutes) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with ab8245 at 5 μg/ml and ab6046 at 1 µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1 hour with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) at 2 μg/ml (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed (ab150088) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labeled 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] - Loading Control (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
    All lanes : 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

    See Abreview

  • Anti-GAPDH antibody [6C5] - Loading Control (ab8245) at 10 µg/ml + Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

    Predicted band size: 40.2 kDa

参考文献

This product has been referenced in:
  • Xu G  et al. Histone deacetylase 3 is associated with gastric cancer cell growth via the miR-454-mediated targeting of CHD5. Int J Mol Med 41:155-163 (2018). Read more (PubMed: 29115379) »
  • Yamada Y  et al. Passenger strand of miR-145-3p acts as a tumor-suppressor by targeting MYO1B in head and neck squamous cell carcinoma. Int J Oncol 52:166-178 (2018). Read more (PubMed: 29115582) »
See all 1110 Publications for this product

レビューと Q&A

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1-10 of 88 Abreviews

Application
Western blot
Sample
Human Cell lysate - whole cell (H1299 cells)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
25 µg
Specification
H1299 cells
Blocking step
Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

投稿 Sep 21 2018

Application
Western blot
Sample
Mouse Tissue lysate - whole (Brain, heart, liver)
Gel Running Conditions
Reduced Denaturing (12% gel)
Loading amount
25 µg
Specification
Brain, heart, liver
Blocking step
Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

投稿 Sep 21 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - other (A549)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
50 µg
Specification
A549
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

投稿 May 22 2018

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

Abcam user community

Verified customer

投稿 Sep 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

Julie Kim

Verified customer

投稿 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

Abcam user community

Verified customer

投稿 May 04 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

Abcam user community

Verified customer

投稿 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

Joey Wu

Verified customer

投稿 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

Francis Djankpa

Verified customer

投稿 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

Mr. Jinwoo Ahn

Verified customer

投稿 May 20 2016

1-10 of 88 Abreviews

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

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