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Does this assay work on frozen mitochondria? Will it work with human PBMC isolated mitochondria?
Asked on Aug 04 2014
The mitotox kits will work with mitochondria from any human tissue. Therefore it will work with mitochondria from human PBMCs.
To isolate mitochondria from human PBMCs I suggest the customer to use ab110170 (Mitochondria Isolation Kit for Cultured Cells) and start with a minimum of 8x10^7 PBMC cells for a yield of 1mg. The mitochondria from PBMCs may be frozen prior to detergent extraction. The higher the mitochondrial preparation protein concentration, the longer the stability at -80ºC storage. The standard mitotox kits are shipped with frozen whole bovine heart mitochondria at 5.5mg/mL. I suggest the customer to do side by side detergent extraction of the bovine heart mitochondria with the PMBC mitochondria and used the BHM mitos in at least 2 or 3 wells of the plate to ensure that the assay is working.
I do not recommend using the MitoTox kits with whole blood.
Answered on Aug 04 2014