Question (62183) | Anti-ABCA1 antibody (ab7360)

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Question

Dear Tineke/Abcam,

We purchased the following Abcam antibodies from Sapphire Biosciences as one single order which were shipped to us as a single lot:

1 AB68466 Rabbit polyclonal to PSAP

1 AB36970 Rabbit polyclonal to Scavenger Receptor BI + BII

1 AB7360 Rabbit polyclonal to ABCA1

1 AB3366 Mouse monoclonal [JSB-1] to P Glycoprotein

1 AB117778 Rabbit polyclonal to Cingulin

1 AB72978 Mouse polyclonal to CNKSR3 Based on our published work these 4 proteins should be expressed in differentiated Caco-2 cells

1 AB111692 Rabbit polyclonal to MAGI3

1 AB83891 Rabbit polyclonal to MPP5


The only one that has worked satisfactorily is AB68466 Rabbit polyclonal to PSAP (see attached file showing these blots). All the others have failed to work as promised. Please note, as previously mentioned, we have used a variety of exposure times with our blots – we have routinely used 10 different exposure times (as the signal increased and also when the signal would have been waning – depending on the results) for each blot using your antibodies to try and optimise what we can visualise with our blots. This has come at some considerable expense in terms of time, developing chemicals and film.

Yes - the other antibodies we tested from our preferred supplier worked. These were purchased at the same time we purchased the Abcam antibodies but they use a different supplier in NZ so were shipped separately. They were tested at the same time we tested yours, using identical conditions and protein samples, and had very low background which is what we have come to expect from their antibodies.

In our previous emails, we filled in questionnaires associated with the failure of AB36970 Rabbit polyclonal to Scavenger Receptor BI + BII, AB7360 Rabbit polyclonal to ABCA1, AB3366 Mouse monoclonal [JSB-1] to P Glycoprotein, and AB117778 Rabbit polyclonal to Cingulin. Despite filling in your questionnaires, you have asked for additional information, which we provided, EXCEPT for the blots as this has taken some considerable time to organise. I have been away at a conference, and as I also work part-time, this has been a pain to organise for you.

During this time we were also evaluating the remaining 4 antibodies and have subsequently found these to be unsatisfactory too. Please see the attached questionnaires for these antibodies. This report of failure to deliver is also within the 6 month time frame of when we received your antibodies.

Do I need to point out, yet again, that we are not stupid. You talked about the samples we tested against (rat, bovine and human Caco-2 cells) and that the antibodies we purchased were not known to cross-react with bovine samples (except MPP5 which was predicted to cross-react). The reason we chose these samples is that although our first set of experiments will ultimately be performed using differentiated Caco-2 cells, some of these targets are highly expressed in liver and we wanted to see if bovine liver could be used as a control – which is why you will note in our blots that we always ran these next to Caco-2 cell extracts to see how they compared. The same goes for the rat colon/rat colon epithelial scrapings – most of the antibodies were known to cross-react with rat or mouse as well as human proteins (also ab36970 is listed as being predicted to cross-react with rat which is not what you said in the email below). Obviously we had good reasons why we wanted to this - we will be doing animal trials in the future amongst other reasons - and again WE ALWAYS COMPARED OUR RESULTS TO OUR HUMAN CACO-2 CELL RESULTS.

I estimate that we have probably spent more money on staff time and other lab consumables trying to get your antibodies to work and fill in your questionnaires, answer yet more questions from you, send you copies of our blots....etc....than it cost us to purchase the antibodies in the first place. This is the first time I have used Abcam antibodies as I usually purchase them from another supplier but they were unable to supply these particular antibodies. My expectation was that if your antibodies performed as expected, I would continue to purchase all my antibodies from you. Given my experience, I will not be purchasing antibodies from you in the future.

Please either replace these antibodies immediately, or better still, please refund our money in full so we can purchase them elsewhere. I don’t want to be bothered with any further questions. You have wasted enough of our time and it’s about time you did the right thing by us.

Answer

Thank you for your reply and all the additional information. This makes the problems you have been experiencing much clearer to understand. It is clear that you have spent a lot of time in trying to optimise these antibodies and I am sorry that they are not providing the expected results.

I am sorry if you feel the questions I have been asking have not been appropriate. The reason I askedfor further information is to try to understand what has been tried already and to try to understand what may be contributing to these problems. With a total of 7 antibodies not working as expected we are taking this very seriously to try and isolate the reason and to offer you a suitable solution as well as preventing this from happening again in the future.

I have been discussing this case with my colleague Dr. Bagrij who has experience in working with a number of the targets you are interested in. I understand the Caco2 lysate you have is very limited and I don't want you to use any more in optimising any of these antibodies. I think it would be worthwhile to try a different lysate to the ones you have been using to see if this makes a difference in the results observed. To this end I would like to send you a pre-prepared Jurkat lysate which have been used as positive controls for the anti-Cingulin antibody (ab117778) and the anti-MPP5 antibody (ab83891). Would you mind trying this lysate with these two antibodies just to see if the expected signal is observed?

You mentioned that you have previously published in regards to the Caco2 cells expressing a number of these targets. Would you mind sharing this reference with me? This information would be very useful in this investigation as well asfor other customers using these antibodies.

Could you also confirm on which date you received the 8 antibodies? I know which order they came from but I want to make sure they reached youwithin the expected time.Were they all delivered in the same package? What was the package like?

I am sorry that you are having these problemsand Iwould like to assure you that I amtrying my best to get to the bottom of this and provide you with a solution.

I look forward to receiving your reply.

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