Anti-Phosphoserine/threonine antibody (ab17464)

Abcam customer care service is really helpful and cooperative. We really
appreciate and give thanks for your help.
I prefer a replacement with a different lot. I will inform you as soon as
the new lot is checked.
Regards

ANSWER:

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 1088417.
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.

as suggested from you in the previous email, a new test of the antibody
(code ab17464) was checked and the phosphorylation of the recombinant
protein was assessed by performing incubation with CHIP.

Answering to your questions, the protein that is being used is AurKA,
commercially available and purified from SF9. The protein is supposed to
be post-translationally modified as a consequence of SF9 cells production.

As you can see from the attached file, the protein is endogenously highly
phosphorylated (revealed by using anti phospho-Thr antibody from Cell
Signaling company) and dephosphorylated in the presence of 40U CHIP.

The protein was incubated in the same kinase buffer used in our previous
experiments and any step was altered.


Since this antibody was used by us in the past and it worked properly, we
can suppose that a mistake in the shipment occured.

Waiting for your istructions

ANSWER:

Thank you for your cooperation in this case.
It may well be that this antibody in vial might have degraded or might have lost the specificity due to unknown reasons. In order to avoid any further delays in your project I am happy to offer you either free of charge replacement with a different lot or a totally different antibody.
Could you let me know how you would like to proceed?

in attaching you can find the filled questionnaire, and the immuno blotting film.
Waiting for instructions from you
My best regards

ANSWER:

Thank you for your email. I am sorry to hear that you are experiencing problems with this antibody.
I have checked the protocol which seems fine to me. However I have doubt about the recombinant proteins you have been using. Could you please give more details about the recombinant protein.
- Have you checked the phosphorylation by any other method?
- What kind of recombinant protein is being used? e.g. human RAS, RAC etc.
- Is it expressed or synthetic?
I also have further suggestion which may improve results.
- I can suggest running whole cell/ tissue lysates to check if the no band is because of antibody or recombinant protein itself?
- Running 10ug will give more prominent results.
I hope these suggestions will help to improve the results. Should the results not improves please do not hesitate to contact me.

I am writing you in reference to our recent order (n. 247 / 2012), of the anti-Phosphoserine / Threonine antibody (cat n. 17464).
Yesterday (May 3th), we tried the antibody.
Unfortunately, after tree tests, we didn't obtain any signal.
For this reason, we would kindly ask for a replacement of the shipment.
Waiting for instructions from you
My best regards.

ANSWER:

Thank you for contacting us.
I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.
I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. 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.

Thanks for your information. It is glad to hear that it is Pan-specific phosphoserine antibody. I am going to use it to detect the phosphorylation of purified recombinant dynein intermediate chain of  mouse origin by a Ser/Thr kinase. Although I am not sure it's serine phosphorylation, it has higher possibility than Thr phosphorylation. Therefore, I am wondering if you could help me check if Abcam has  pan-specific phospho-Ser/Thr  antibody. Otherwise, I am going to get  this Pan-specific phosphoserine antibody and pan-specific phosphothreonine antibody(if there is any). if our research is qualified for discount, please let me know. Thanks

ANSWER:

We do offer two products which may meet your needs, ab17464 and ab117253.

While not truly Pan phospho Ser/ThrAb14764 detects phosphoserine or phosphothreonine in the context of tyrosine, tryptophan or phenylalanine at the -1 position or phenylalanine at the +1 position. This product has been tested and guaranteed to work inICC/IF,ELISA,IP,and WB and has been referenced in seven publications to our knowledge. The product page is:

http://www.abcam.com/Phosphoserine-threonine-antibody-ab17464.html

Ab117253detects free and conjugated Phosphoserine/threonine as well as their modified cellular proteins. It does not cross-react with nonphosphorylated tyrosine, serine, threonine, AMP, or ATP. It has been tested and guaranteed to work inELISA,Dot Blot on human tissues. Again, I would like you to be aware that for each antibody of this type certain proteins known to contain phosphorylated serine and threonine may not be detected due to steric hindrance.The product page for this is:

http://www.abcam.com/Phosphoserine-threonine-antibody-ab117253.html

For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it works:

The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody at the full price (100%) of the antibody you have tested. The terms and conditions applicable to this offer can be found here: http://www.abcam.com/collaborationdiscount.

To determine if one of these products would qualify for participation in our 100% Abreview testing discount program could you tell me more about the application and species you would be testing the product on?

I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.