Anti-Scavenger Receptor BI + BII 抗体 (ab36970)

Thank you for taking the time to contact us. I am sorry to hear you have some concerns regarding this antibody.

In this case, I would suggest the white powder you are seeing may be precipitate. We recommend that aliquots of antibody should be no smaller than 10 µl where possible. The smaller the aliquot, the more the stock concentration is affected by evaporation and adsorption of the antibody onto the surface of the storage vial. This evaporation could also result in precipitation, particularly if the antibody is stored frozen for a long period, such as a year.

I can recommend to mix the precipitate very well using a pipette, and try vortexing as well. I hope this would provide a more homogenous solution for you to try in your experiments.

I would like to reassure you that this antibody is tested and covered by our 6 month guarantee for WB, ICC/IF, IP, blocking and in mouse and human samples. We can also guarantee the general quality of the antibody for 6 months. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund. In this case, I am sorry to confirm that as the antibody was purchased over 6 month ago it will regrettably no longer be covered by our 6 month guarantee on this occasion.

Please do not hesitate to let me know if you have any further questions or concerns.

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.

Thank you for your enquiry

We obtain this product from an external source and unfortunately they are not able to provide specific information about the sample preparation.

I reviewed the literature and found the following protocol that you may find useful:

1. Used 1-2 liver lobes.
2. Homogenize tissue in PBS/Protease Inhibitor cocktail.
3. After centrifugation resuspend the pellet in approx. 400µl RIPA lysis buffer 30min (vortex every 5-10 min).
4. Centrifuge to give whole cell extracts.
5. To improve the amount of sample extracted, repeat this lysis step with the pellet from the centrifugation and pool the supernatant of the 1st and the 2nd lysis.

We also stock a range of liver lysates:

Liver (Mouse) Whole Cell Lysate - normal tissue, 0 days old
www.abcam.com/ab7264

Liver (Mouse) Whole Cell Lysate - normal tissue, 7 days old
www.abcam.com/ab7266

Liver (Mouse) Whole Cell Lysate - normal tissue
www.abcam.com/ab29301

Liver (Human) Tissue Lysate - adult normal tissue
www.abcam.com/ab29889

Please let me know if there is any more information you need.