Question (59074) | Anti-ABCA1 antibody (ab7360)

Go to datasheet (ab7360)

Question

Please see below for the response from the customer;

I’m very frustrated by Abcam’s response. It took a long time to fill in the questionnaires for each of the antibodies and some of their additional questions were already answered in the questionnaire (Q3 and Q6). Q6 is quite insulting, even more so when I had specifically answered that I’d used anti-mouse secondary antibodies.

It also took them 4-5 working days to reply to the questionnaire answers, now they want more information, taking more of my time, and I guess I have to wait another 4-5 working days for their next response.

Quite frankly, this is unacceptable.

In short,

Q1. the same blocking agent was used consistently throughout the western experiment. So, if BSA was used for blocking, it was also used for the primary and the secondary. When it didn’t work, we tried milk etc.

Q2. For most of the repeats, blots weren’t stripped. We made ran fresh gels and blotted these (exactly the same loading concentrations and conditions). Therefore, stripping was not the issue. ALWAYS, we ran another gel with the same loading concentration which was stained with coomassie. ALWAYS we ran coloured markers so we could check that the protein had transferred. We have never had trouble with transfer using the i-blot.

Q3. Please read the answers I have already supplied.

Q4. Exposure times varied from 15 s through to 15 min. We also used 2 pieces of film so that the bottom film acted as a filter. We developed at least 8 pieces of film for each blot each time we repeated it and also used a range of exposure times. Faint blots were left longer while those with high background were just left for a few seconds. High background blots were also re-exposed after 1-2 hours as the signal was waning. We covered the entire range.

Q5. This will take a lot of time as we’ve done lots of blots. They will have to insist this is absolutely necessary then pay for my time. It’s their antibodies that aren’t working so I shouldn’t have to defend my results. My time will probably cost more than their antibodies do to replace.

Q6. The secondary antibodies were brand new and worked because, as previously answered, we had magic markers which only show up if the secondary antibody is working.

My expectation is that Abcam will hurry up and come to the party with new, replacement antibodies in a timely manner and stop wasting our time. If they don’t, we won’t be using them again and as we work for a large research institution with a large international network of collaborators, won’t be recommending them to anyone.

Answer

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.

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