Hi everyone.
I am considering buying the SIRT1 Activity fluorometric Assay Kit from Abcam (cat. no. ab156065). However, after reading very carefully the protocol booklet, I am quite dubious. What I am looking for is a method that detects ONLY the activity of SIRT1. I have the feeling that the kit tries to eliminate the contribution of other de-acetylating sirtuins by measuring sirtuin-dependent deacetylase activity on proteins derived from isolated nuclei. Such an approach would ignore the fact that extranuclear SIRT1 is active on cytoplasmic proteins. Moreover, how can the supplier be sure that the kit does not detect the activity of other nuclear sirtuins (e.g., SIRT2/6/7)? Unfortunately, the datasheet provides no clear information with regard to the substrate peptide used. Does the kit use a substrate peptide that is very specific for SIRT1? If yes, does anybody know which one? Has anyone ever used the Abcam's SIRT1 Activity fluorometric Assay Kit?
Obviously, I am waiting for a reply from the Abcam's technical support.
Thanks in advance.
Stefano Falone Department of Life, Health and Environmental Sciences
University of L'Aquila
Email [email protected]
..."Obviously, I am waiting for a reply from the Abcam's technical support."
Abcam's reply:
"Our scientific support team is currently working on your inquiry. We need to contact other scientists to help with your query and we will reply to you as soon as we have further information."
I just received the official reply from Abcam's technical support. Speechless.
"...after communications with our laboratory, I have received the following answers to your questions:
They have confirmed that the activity of other sirtuins cannot be excluded by measuring sirtuin-dependent deacetylase activity on proteins derived from isolated nuclei. The substrate provided with the kit is deacetylated not only by sirt1, but also by recombinant sirt3 but we have not tested yet the "native" SIRT3 with the kit."
The kit's commercial name is "SIRT1 Activity Assay Kit"! How can they sell a kit with such a name if they are aware that the substrate provided with the kit is deacetylated also by other sirtuins?
Bad day for Abcam's customers.
Stefano
I'm thinking of using the Abcam Adenosine Assay Kit, but am hesitant considering your experiences with the Abcam kits.
Please advise.
Dear Hilja Eelu,
we are currently setting the experiment. I'll inform you on the results obtained.
Regards
Stefano
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