Anti-GAPDH 抗体 [mAbcam 9484] - Loading Control (ab9484)

Reviews (60)Q&A (7)References (542)

製品の概要

  • 製品名

    Anti-GAPDH antibody [mAbcam 9484] - Loading Control
    GAPDH 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control

  • 由来種

    Mouse

  • 特異性

    In western blot, this product typically gives a lower signal in rat lysates compared to human and mouse lysates.

  • アプリケーション

    適用あり: WB, IHC-P, Flow Cytmore details

  • 種交差性

    交差種: Mouse, Rat, Rabbit, Chicken, Cow, Dog, Human, Pig, Xenopus laevis, Cynomolgus monkey, Chinese hamster

  • 免疫原

    Full length native protein (purified) corresponding to Human GAPDH.

  • 特記事項

    For Western blotting, do not use milk for blocking. Our labs have extensively tested the blocking conditions for this antibody and recommend using 5% BSA for 1 hour. The comparison data is shown in the images section. 

    This antibody clone [mAbcam 9484] is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab9484 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
WB Use a concentration of 0.1 - 1 µg/ml. Predicted molecular weight: 36 kDa.

Do not block with milk. Block with 5% BSA for 1 hour. Our labs have thoroughly investigated the blocking conditions for this antibody. We found that milk significantly decreases the signal and is therefore not a suitable blocking agent for this antibody (see images).
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

We recommend Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879) secondary antibody

ターゲット情報

画像

  • Western blot - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)
    Western blot - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)
    All lanes : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 1 µg/ml

    Lane 1 : HeLa Whole Cell Lysate
    Lane 2 : NIH 3T3 Whole Cell Lysate
    Lane 3 : PC12 Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 36 kDa


    Exposure time: 1 minute


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab9484 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406

  • Western blot - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)
    Western blot - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)
    All lanes : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 1/5000 dilution

    Lane 1 : Hela whole cell (Human)
    Lane 2 : 3T3 cell (Mouse)
    Lane 3 : Rat brain
    Lane 4 : Xenopus embryo
    Lane 5 : Chicken Liver
    Lane 6 : EBTr cell (Cow)
    Lane 7 : CHO cell (Chinese hamster)
    Lane 8 : Pig liver

    Secondary
    All lanes : Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size: 36 kDa
    Observed band size: 40 kDa
    why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    The membrane was blocked in 5% BSA in TBST for 1 hour, then incubated for 1 hour in primary antibody diluted in TBST.

  • Western blot - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)
    Western blot - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)
    Lanes 1-5 : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 1/1000 dilution (Blocked in 5% milk)
    Lanes 6-10 : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 1/1000 dilution (Blocked in 5% BSA)

    Lanes 1 & 6 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
    Lanes 2 & 7 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lanes 3 & 8 : A431 whole cell lysate (ab7909)
    Lanes 4 & 9 : Jurkat whole cell lysate (ab7899)
    Lanes 5 & 10 : HEK293 whole cell lysate (ab7902)

    Lysates/proteins at 20 µg per lane.

    Secondary
    Lanes 1-5 : Goat anti-Mouse (HRP conjugated) at 1/5000 dilution
    Lanes 6-10 : Goat anti-Mouse (HRP conjugated) at 1/5000 dilution

    Predicted band size: 36 kDa
    Observed band size: 40 kDa why is the actual band size different from the predicted?



    The membrane 1-5 was blocked in 5% milk (1 hour). The membrane 6-10 was blocked in 5% BSA (1 hour). Milk is not a suitable blocking agent and significantly decreases the signal on the membrane.

  • Western blot - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)
    Western blot - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)
    Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 0.5 µg/ml + HeLa cell lysate

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) (ab6789) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under non-reducing conditions.

    Predicted band size: 36 kDa


    Exposure time: 30 seconds
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)
    IHC image of GAPDH staining in human liver FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab9484, 5µg/ml, for 8 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  • Flow Cytometry - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)
    Flow Cytometry - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)

    Overlay histogram showing HeLa cells stained with ab9484 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab9484, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was goat anti-mouse DyLight® 488 (IgG H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

参考文献

This product has been referenced in:

  • Gordon D  et al. Single-copy expression of an amyotrophic lateral sclerosis-linked TDP-43 mutation (M337V) in BAC transgenic mice leads to altered stress granule dynamics and progressive motor dysfunction. Neurobiol Dis 121:148-162 (2019). Read more (PubMed: 30290270) »
  • Mulati K  et al. VISTA expressed in tumour cells regulates T cell function. Br J Cancer 120:115-127 (2019). Read more (PubMed: 30382166) »
See all 542 Publications for this product

ab9484 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

レビューと Q&A

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1-10 of 67 Abreviews or Q&A

Western blot abreview for Anti-GAPDH antibody [mAbcam 9484] - Loading Control

 Good
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (liver, HepG2)
Gel Running Conditions
Reduced Denaturing
Loading amount
10 µg
Specification
liver, HepG2
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Read More

Abcam user community

Verified customer

投稿 Dec 04 2017

Western blot abreview for Anti-GAPDH antibody [mAbcam 9484] - Loading Control

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Rat Tissue lysate - whole (Retina)
Gel Running Conditions
Reduced Denaturing (12% gel)
Loading amount
20 µg
Specification
Retina
Blocking step
Licor Odyssey Blocking Buffer as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 4°C
Read More

Abcam user community

Verified customer

投稿 Aug 01 2017

Western blot abreview for Anti-GAPDH antibody [mAbcam 9484] - Loading Control

 Inconclusive
Abreviews
abreview image
Application
Western blot
Sample
Zebrafish Tissue lysate - whole (Liver, RPE, retina)
Gel Running Conditions
Reduced Denaturing (12% gel)
Loading amount
20 µg
Specification
Liver, RPE, retina
Blocking step
Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Read More

Abcam user community

Verified customer

投稿 Mar 24 2017

Western blot abreview for Anti-GAPDH antibody [mAbcam 9484] - Loading Control

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

Abcam user community

Verified customer

投稿 Mar 17 2017

Western blot abreview for Anti-GAPDH antibody [mAbcam 9484] - Loading Control

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (Liver)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
20 µg
Specification
Liver
Blocking step
Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Read More

Abcam user community

Verified customer

投稿 Dec 14 2016

Western blot abreview for Anti-GAPDH antibody [mAbcam 9484] - Loading Control

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Rat Tissue lysate - whole (muscle)
Gel Running Conditions
Reduced Denaturing (9)
Loading amount
50 µg
Specification
muscle
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More

Mr. Baptiste Jude

Verified customer

投稿 Oct 07 2016

Immunocytochemistry/ Immunofluorescence abreview for Anti-GAPDH antibody [mAbcam 9484] - Loading Control

 Excellent
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (H1299 Lung cancer cell line)
Permeabilization
Yes - 0.1% v/v TritonX-100
Specification
H1299 Lung cancer cell line
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde
Read More

Dr. Dimitra Kalamida

Verified customer

投稿 Jun 06 2016

Western blot abreview for Anti-GAPDH antibody [mAbcam 9484] - Loading Control

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (PERIPHERAL BLOOD MONONUCLEAR CELLS)
Gel Running Conditions
Reduced Denaturing (12% SDS PAGE)
Loading amount
30 µg
Specification
PERIPHERAL BLOOD MONONUCLEAR CELLS
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Mr. Siddharth Mehra

Verified customer

投稿 Jun 02 2016

Western blot abreview for Anti-GAPDH antibody [mAbcam 9484] - Loading Control

 Excellent
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

投稿 Mar 17 2016

Western blot abreview for Anti-GAPDH antibody [mAbcam 9484] - Loading Control

 Good
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse hepatocytes)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
mouse hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Read More

Abcam user community

Verified customer

投稿 Mar 14 2016

1-10 of 67 Abreviews or Q&A

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