Overview

  • Product name
    Anti-GAPDH antibody [mAbcam 9484] - Loading Control (HRP)
    See all GAPDH primary antibodies
  • Description
    Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (HRP)
  • Host species
    Mouse
  • Conjugation
    HRP
  • Tested applications
    Suitable for: WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Chicken, Cow, Dog, Human, Xenopus laevis, Chinese hamster
  • Immunogen

    Full length native protein from human erythrocytes.

  • Positive control
    • WB: HeLa, HEK-293, NIH/3T3 and PC-12 whole cell lysates.
  • General notes

    According to our customer's feedback this antibody does not recognise meningococcal GapA1 or GapA2 (GAPDH) recombinant proteins.

    This antibody has been conjugated with HRP. The concentration of the original antibody was 1 mg/ml, following conjugation the concentration observed will be higher.

Properties

Applications

Our Abpromise guarantee covers the use of ab9482 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
WB 1/1000 - 1/5000. Detects a band of approximately 40 kDa (predicted molecular weight: 40.2 kDa).

NOT SUITABLE for blocking with milk. Block in 5% BSA for 1 hour. Our labs have thoroughly investigated the blocking conditions for this ab following concerning customer feedback on the lack of signal with some vials. We found that milk significantly decreases signal and is therefore not a suitable blocking agent for this ab (see images of our tests on the datasheet of the unconjugated version, ab9484). This change in characteristics of the ab is due to a recent update to the production process.

Target

Images

  • All lanes : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (HRP) (ab9482) at 1/1000 dilution

    Lane 1 : HeLa Whole Cell Lysate
    Lane 2 : Hek293 Whole Cell Lysate
    Lane 3 : NIH 3T3 Whole Cel lLysate
    Lane 4 : PC12 Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Developed using the ECL technique.

    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?


    Exposure time: 4 minutes

References

This product has been referenced in:
  • Chang HC  et al. Bhlhe40 differentially regulates the function and number of peroxisomes and mitochondria in myogenic cells. Redox Biol 20:321-333 (2019). Read more (PubMed: 30391825) »
  • Fang S  et al. Osteogenic effect of bone marrow mesenchymal stem cell-derived exosomes on steroid-induced osteonecrosis of the femoral head. Drug Des Devel Ther 13:45-55 (2019). Read more (PubMed: 30587927) »
See all 118 Publications for this product

Customer reviews and Q&As

1-3 of 3 Q&A

Question

You contacted Susan Lindsay regarding the GAPDH antibody Abcam reference 163989, purchase order 250103139. Susan ordered the antibody but it was actually myself that used it. I actually found that the antibody did not work at all. I used it as a loading control for CHO-K1 cells. After probing the membrane with the antibody for my specific protein, the membrane was stripped and prepared as detailed in the ECL-Plus Kit (Amersham) before using the HRP-GAPDH antibody which was incubated at a 1:5000 for 1hr at room temperature then washed in PBST and detected using the ECL detection kit. The film showed absolutely no bands on it at all. I was worried I had done something wrong so I stripped the membrane again and reprobed it with our Abcam beta actin mouse monoclonal antibody then used a secondary biotinylated mouse antibody, streptavidin then ECL which gave very strong beta actin bands. I was surprised that the HRP GAPDH did not work as I have previously used your non-conjugated GAPDH antibody after stripping and it has worked fine. I only bought the HRP one because I thought it would save time by missing out some of the incubation steps. After discussing the problem with other colleagues it appears that I am not the only person experiencing problems with this antibody. Another lab colleague Mark Moore, also said that the antibody did not work for him either. You can contact him to get his feedback also. I hope this information is helpful. If it possible, a full refund would be appreciated.

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Answer

Many thanks for your valuable feedback on your recent experience of ab9482. In light of your feedback and that of several other customers we have removed all stock of this antibody and are offering our apologies for this unusual problem and a refund or replacement vial of another antibody. We have found that some customers are having difficulty with this product, while others are delighted with the same lot, we are very puzzled as to the reason for this and are investigating this extremely seriously in the laboratory. The quality of our antibodies is of utmost importance to us and we are very concerned about this case. We have contacted all customers who recently purchased this antibody so I hope that Mark received our e-mail. Please do let him know that we will offer him a refund or replacement if he has not received our e-mail. A refund is being arranged with our accounts department. Once again please accept our apologies for this unfortunate rare event, I hope this will not deter you from purchasing from Abcam in the future.

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Answer

Thank you for your phone enquiry earlier this morning. I have had a look at the records for this antibody (Ab9482) and we have experienced some mild batch variation. It has now been extensively tested in the laboratory again, giving good results but at a higher concentration. I would recommend you repeat the western blot at eg 1:1000 or even 1:500. I hope this helps. I have placed a link to the questionaire here in case you have any more problems. Please do not hesitate to send it back if you are still unable to obtain satisfactory results, and I will be happy to help. https://www.abcam.com/index.html?section=western&pageconfig=technical&mode=questionaire Good luck! With thanks.

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Question
Answer

Thank you for your enquiry. The only difference between these two antibodies is that ab9482 is the HRP-conjugated version of ab9484. The advantage of ab9482 being HRP-conjugated is that there is no need for a secondary antibody.

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