Question (76335) | SIRT3 Activity Assay Kit (Fluorometric) (ab156067)

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Question


1) How specific is the assay? There are two other SIRT proteins in the mitochondria, and both are NAD dependent. It seems as if this assay will measure activity of all of these proteins, no?
2) What is the difference between the mitochondrial isolation kit and using a normal Triton/Digitonin protocol for mitochondrial extraction?
3) Are intact mitochondria required for the SIRT3 activity assay, or are mitochondrial extracts enough?
4) Would it be possible to receive a test sample of either of these kits, to see if they work for what I want to do before buying the whole kit?

Answer

1) How specific is the assay? There are two other SIRT proteins in the mitochondria, and both are NAD dependent. It seems as if this assay will measure activity of all of these proteins, no?
I assume you are referring to SIRT5 as the second SIRT protein? We know that SIRT1 (recombinant) can catalyze deacetylation from the substrate of SIRT3 Activity Assay Kit (Fluorometric) and SIRT2 cannot. We don't know about SIRT5.

2) What is the difference between the mitochondrial isolation kit and using a normal Triton/Digitonin protocol for mitochondrial extraction?
The kit comes with optimized detergent concentrations to isolate mitochondria and an optimized isosmotic buffer to reduce swelling.

3) Are intact mitochondria required for the SIRT3 activity assay, or are mitochondrial extracts enough?
We haven't used SIRT3 Activity Assay Kit (Fluorometric) with native SIRT3 samples ourselves.

4) Would it be possible to receive a test sample of either of these kits, to see if they work for what I want to do before buying the whole kit?
Unfortunately, we do not offer test samples of our products due to logistical issues. However, all our products are covered by our 1 year Abpromise guarantee which states that if the product does not work as stated on the datasheet, we will replace or refund the product.

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