Anti-53BP1 antibody (ab21083)

Thank you very much for your reply.

That is no problem to arrange. I have organised a new order for you for a new vial of ab21083. This is on the order number xxxx(Purchase order numberxxxxx). Please let me know if you have any problems receiving this.

I hope this one performs better for you. However, if you do continue to have problems please do let me know.

Until then, I wish you all the best with your research.

Thank you for taking time to complete our questionnaire. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. I cannot explain why the activity of the antibody diminished over the time you had it.

One thing I would check is that you do not have a self-defrosting freezer. These automatically defrost, raising and lowering the temperature, forcing the antibody through freeze-thaw cycles, potentially decreasing its activity.

Having reviewed the protocol details, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial. There is only one thing I would point out which may be worth taking into consideration. You have used 0.5% TritonX in the antibody diluent as well as the wash buffers. This as well as the permiabilisation step may be too strong. I would use one 10 minute permiabilisation step (using 0.2% Triton X in PBS) at room temperature. Following this I would only use 0.1% Tween 20 in your diluent buffers. You did not mention explicitly but are you using PBS or TBS for the diluent buffers?

If you would like, I can send you a new vial of ab21083 to be sent to you to see if this makes a difference in your staining? If you would like for me to arrange this please could you confirm I have the correct addressbr/>Alternatively, if you would prefer I can offer a credit note, or refund in compensation.

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

Thank you for contacting us yesterday and sorry for the delay in getting back to you.

I am sorry to hear that you have been having some difficulty in using the Anti-53BP1 antibody (ab21083). We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

As we discussed on the phone, I am attaching our questionnaire so that I can gather further information regarding the samples tested and the protocol used. Once I have receive the completed questionnaire, I will look at the protocol and see if there are any suggestions I can make that may improve the results.

If you could include some images of the staining when the antibody worked and how the latest experiments have been that would be very helpful.

This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

I look forward to receiving your reply.

Thank you for your inquiry.

Unfortunately, we are not able to release the immunogen sequence for this particular antibody as it is proprietary information. However, I can tell you that it falls within the carboxy-terminus of h53BP1.

We aim to provide as much information as possible to our customers, so I am sorry that this has not been possible on this occasion.

Should you have any further questions, please do not hesitate to contact us.

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hearthe customer has haddifficulty obtaining satisfactory results from this antibody.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

As far as I know, Biomoles have been made aware that they are not an authorized distributor of Abcam products. Regrettably, I am sorry this means we are not able to provide replacements or credit notes for faulty items purchased and resold by Biomoles. However, we are pleased to help as much as we can with technical support. Reviewing this case, I would like to offer some suggestions to help optimize the results. I would also appreciate if you can confirm some further details:

1. I recommend to check the amount of protein loaded. Try a protein assay on the sample and load 20 -30 ug of total protein per lane of the gel to ensure there is enough protein for detection.

2. Please confirm what type of cancer cells are being tested? What positive controls have been used? For example, HeLa whole cell lysate would be a suitable positive control.

3. Could you confirm if the transfer of protein to the membrane and quality of the sample has been assessed using a loading control?

4. Please confirm what dilutions of antibody have been tried? This may require some optimization.

5. Is the secondary antibody working well with other antibodies?

I hope this information is helpful. Thank you for your cooperation and for your understanding of our policy regarding unofficial distributors. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

Thank you for your enquiry. We do notice that there is some size discrepancy between the calculated MW vs. observed MW with the ab21083 antibody. This product has been tested in house on Hela cell lysate and the antigen that was used to generate the antibody by ELISA and WB. We also found that the peptide blocks antibody detection by WB. Based on our testing and data collected we concluded and feel confident that the antibody is detecting the 53BP1 protein. I have included PubMed IDs for several publications which describe detection of the 53BP1 protein by WB. The literatures indicate that 53BP1 protein migrates significantly slower than its predicted size. PubMed ID: 11238909 11134068 9748285 15840649 This product is one of our top selling antibodies and we have not received complaints regarding detection of the incorrect protein. This is currently all the information we have available. But if we do happen to obtain more data, we will be sure to forward you the information.