Anti-SIRT1 antibody (ab12193)

            Dear Abcam, i would need a good sirt1 antibody to use in WB and IHC for rat cell lines( PC12). Does the ab12193 work on rat? Otherwise, could you help me finding the right ab? Many Thanks,

ANSWER:

Thank you for contacting us. Ab12193 has not been tested in rat, neither with IHC. If you wish to test it in rat, or with IHC, I can offer a discount code to test it as part of our antibody characterisation plan. Therefore, I can offer a discount off a future purchase if you buy ab12193 now, test it in Human / IHC and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody.  However, there are other antibodies in our catalogue against the same protein that have been tested in rat samples and characterised by WB. If you wish to test them with IHC, you can do it under our Abreview system. Please have a look at their datasheets: http://www.abcam.com/SIRT1-antibody-19A7-ab110304.html http://www.abcam.com/SIRT1-antibody-ab75435.html To find out more about our Abreview system, please visit:  http://www.abcam.com/abreviews I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

It states that this is a SIRT1 antibody but the immunogen is stated as being "Synthetic peptide, corresponding to C-terminal amino acids 722-737 (with C-terminal added lysine) of Mouse SIR2/SIRT2."

Please clarify.

ANSWER:

Thank you for your enquiry. That was a typo in the immunogen section for this antibody. The correct statement is "Synthetic peptide, corresponding to C-terminal amino acids 722-737 (with C-terminal added lysine) of Mouse SIR2/SIRT1. This sequence is 62% homologous to the corresponding human sequence."

I have updated the online datasheet and apologize for any confusion.

So far this antibody has been tested for cross-reactivity with mouse and has not yet been tested for cross-reactivity with any other species. I did not see the rat sequence in the SwissProt database, the mouse and human sequences are there. As stated above, there is 62% homology between the peptide and the human SIR2 sequence.

Should you decide to go ahead and purchase this product, please let us know how you get on by submitting an Abreview and in return we will award you 50 Abcam Points, which can be redeemed on a number of rewards (a further 100 points will be offered for an image).

Please let me know if you require any further assistance.

I have 2 questions about the antibody against mouse sirtuin 2 (ab12193): As far as I (and the swissprot database) know, the protein has a molecular weight of 43 kDa. How come that the antibody detects a band at 110 kDa? And as the protein is localized in the cytoplasm, why is a nuclear extract described as positive control and the protein described as important for histone research?

We are very interested in an antibody against mouse sirtuin 2, I would just like to make sure that it really is against the protein that I'm thinking of.

Thanks a lot in advane for an answer,

ANSWER:

I am sorry for the delayed response to your enquiry. In the meantime, we have updated the datasheet for this Rabbit polyclonal antibody to SIR2 / SIRT2. The predicted molecular weight is indeed 43 kDa, 110 kDa was a mistake. The fact that we described the location as cytomplasmic and yet recommended a nuclear extract as a positive control was also confusing. We have done some more research and in light of our findings have added the following to the datasheet to describe the cellular localisation: "Cytoplasmic for short periods of time until complexes form with Sir2, Sir3 and Sir4. The complexes are recruited to their chromosome targets (nuclear) via interactions with DNA-binding proteins, followed by deacetylation of histones H3 and H4. Therefore, this protein should be easily detectable in the nuclei and nuclear extracts.

Thank you for bringing these matters to our attention.