Question (76710) | SIRT3 Activity Assay Kit (Fluorometric) (ab156067)

Go to datasheet (ab156067)

Question

I would like to use this kit (ab156067 sirt3 assay kit), in order to measure endogenous SIRT3 activity in cell. According to the cover sleeve the kit is intended " For the quantitative measurement of SIRT3 activity in cell lysates".
However, after reading the booklet, i understand that is not possible to measure endogenous cellular SIRT3 activity with this kit, which only detects activity of recombinant or immunoprecipitated SIRT3 protein.
Is that right? Can you confirm my question?

Answer



I am happy to confirm that crude cell lysate can be used as samples with this kit but as this often needs a lot of optimisation we would recommend to prepare mitochondria and use a mitochondrial lysate.

I can recommend to use one of our mitochondria isolation kits for this purpose:

ab110171 (Mitochondria Isolation Kit for Cultured Cells (with Dounce Homogenizer))

https://www.abcam.com/index.html?datasheet=110171 (or use the following: https://www.abcam.com/index.html?datasheet=110171).

ab110169 (Mitochondria Isolation Kit for Tissue (with Dounce Homogenizer))

https://www.abcam.com/index.html?datasheet=110168 (or use the following: https://www.abcam.com/index.html?datasheet=110169).

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