Anti-ABCA1 antibody (ab7360)

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued an alternative free of charge replacement of ab7360 with the order number 1214026.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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.

Thank you for your reply.

I am very sorry that you have not been satisfied with the way we have handeled your complaints. I do understand and appreciate the amount of time you have spent it the laboratory. Our policy is that we are happy to offer a refund, credit note or free of charge replacement when a product is not working in a successfully tested applications or species (and the product has been purchased in the last 180 days).

However, we often find that suggesting some scientifically thought out optimisation tips improves the results. This is obviously a preferable outcome.So I hope you can understand that our policy is to offer the best technical support possible to help optimise the results first.

It is also important for us to understand what is contributing to the problems encountered, as if there is a problem with the antibody, forexample with a particular lot, we need to try to find out why this has happened and if any other customers may be affected and if we need to let them know.

I am sure you can understand that with four antibodies not providing satisfactory results for you we need to take it especially seriously, I do not want to simply offer replacement for them to perform in exactly the same way and further waste your time.

I am sorry that I did not read the information regarding the anti-mouse antibody used with ab3366 properly. This was an error on my part and I apologize. The reason I wanted to have a look at the images of the blots was to try to ascertain where the high background may be coming from. With the antibodies used, none have been tested with bovine samples so we would not be able to know if they would cross react or not. Similarly, ab7360 and ab36970 have not been tested with rat samples as yet, although they would be predicted to cross-react due to their sequence homology. That leaves onlytheCaco-2 cell samples for ab7360 and ab36970. It would be very helpful to understand the results observed more fully if you would be able to provide a few representative images of the blots obtained.

You have mentioned that other antibodies have been used with the samples and have performed as expected. Are any of these from Abcam, or are they from other companies? Were any of them delivered in the same shipment as the 4 antibodies which are not working? I am trying to ascertain if there may have been a problem with your particular delivery which affected the antibody activity. This is important as with some of the antibodies involved, we only have the same lots available as you have already tried and I need to ascertain if this is worth trying again for you.

I am sorry for the delay youhave experienced in our handling of your complaints. I am sorry for this and the inconvenience caused. I will try to resolve the situation as quickly as I can.

Thank you for reporting the problems encountered with these antibodies and for taking the time to complete the questionnaire. I am sorry to hear that these antibodieshave not providing satisfactory results so far.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

Having reviewed this case, there are a few further details I would like clarified if possible:

1. You have mentioned that BSA, milk and Roti block were tried. When each blocking agent was tried, was the same blocking used in the primary antibody diluent? For example, when using the BSA blokcing, did you use BSA in the antibody diluent or did you continue with milk for this? Was this the case for the secondary antibody as well?

2. Could you explain how the membranes were stripped between each staining?

3. Were the same transfer conditions used for each antibody?

4. What exposure times were used?

5. Could youshare some images of the blots obtained? With each lane labelled with the samples and the molecular weight markers. If its not too much trouble, could the images of the blotting with the xxxxxx antibody and the acting loading control also be included. This would help greatly in understanding more fully the problems encountered with the antibodies.

6. With the anti-P Glycoprotein antibody (ab3366), did you use the same secondary antibody as with the other antibodies? Please bear in mind that this is a mouse monoclonal antibody and will not be detected by the anti-rabbit secondary antibody. Was a different antibody used? If so, has this one been used successfully previously?

In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

I look forward to receiving your reply.

I'm sorry to hear you are having a problem with ab7360. I would like to suggest the following modifications to your protocol: The recommended dilution range for this antibody is 1/500 - 1/1000. You may be using it at too high a dilution. I did not see your incubation conditions with this antibody from your enquiry. Since you are seeing no signal, I would suggest removing the milk from the primary antibody solution and incubating at 4C overnight. The milk in the primary antibody solution can overblock the membrane, preventing the antibody from binding to its target. I would continue incubating the sample in loading buffer for 30 minutes at 37C. This protein can form aggregates, preventing migration into the gel matrix. For how long did you block? If you have blocked for longer than one hour, you may wish to reduce this step to 1 hour, or even 30 minutes. Did you check (e.g. Ponceau stain) to see that the transfer was efficient? Please let me know if this helps and do not hesitate to contact us for further advice.