Anti-Rad17 (phospho S645)抗体(ab3620)

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 a free of charge replacement with the order number 1048217 (ab63442).

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.

I am sorry to hear that the results does not improve. I can offer you another antibody we have following antibodies against the same target

ab63442; https://www.abcam.com/Rad17-phospho-S645-antibody-ab63442.html

ab115876; https://www.abcam.com/Rad17-phospho-S645-antibody-ab115876.html

Please check ab115876 is a mouse polyclonal antibody, you would need anti mouse secondary if you opt for this.

Let me know which ab you would like to try.

Well that's true. BSA reduces the background only, it does not have any effect on ab specificity.

Anyway to proceed further in this case, I would like to ask if you are happy in trying a different lot or a different antibody. Alternatively I can also provide you full refund or credit note, which you can use for future purchases. Let me know the option you would like to go for.

Many thanks for your cooperation in this case, I highly appreciate that.

Thank you very much for your email.

We use 5% BSA (Sigma) in TBST for blocking and for diluting secondary and primaries. Milk in not recommended for phospho specific antibodies because milk contains casein which is a phosphoprotein; This is why it causes high background because the phospho-specific antibody detects the casein present in the milk. Please be advised that BSA will only reduce the background i.e. membrane dirtiness, it will not effect the antibody specificity. So if the membrane shows high background then I would certainly recommend trying BSA.

The other important point is, whether the cells naturally express the protein or not. I am sorry the info of cell line wasn't included in your email so I can't be sure about it. I would however recommend using Hela lysates as positive control. Are you sing human cell line?
Actin or GAPDH can be used as loading control. Have you tried any of these?

I hope these suggestions will help to improve your results. If the results do not improve please do not hesitate to contact me.

Thank you for your enquiry regarding ab3620 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.

The protocol looks fine to me however I would like to make few comments/suggestions that might help to improve your results and that you could consider trying:

ab3620 is a phospho specific antibody. We used 10Gy IR treated Hela cells to test the specificity of this antibody. I would recommend treating your sample cells there by inducing the phosphorylatuion of amino acid chemically or by radiations.

Milk is not recommended as blocking reagent with phospho specific antibodies. Please use BSA.

Please try both of these suggestions and let us know the results; we will be happy to help further.

I am sorry to hear that you have been experiencing problems using this product in the application that you wish.



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Please could you provide some further details of the protocol used and complete the following form (attached as a word document). It would be much appreciated if you could attach an image to the response.



Thank you for your understanding and co-operation in this matter. I look forward to hearing from you soon and resolving this issue as soon as possible.

To our knowledge, these three antibodies have not been used in parallel, which would be required to determine if there are truely differences in the apparant molecular weight of the protein recognized by each antibody. The differences in the reported apparant molecular weights of the proteins detected by the 3 antibodies is likely owing to differences in molecular weight markers that where used, position of the sample relative to the positon of the molecular weight markers in the gel, and size and composition of the resolving gels. The difference in relative mobility of the protein recognized by the goat anti-Rad17 and the phospho specific antibody is certainly within experimental error.