Skip to main content
. Author manuscript; available in PMC: 2014 Sep 1.
Published in final edited form as: Exp Gerontol. 2013 Jun 24;48(9):947–959. doi: 10.1016/j.exger.2013.06.002

Fig. 1.

Fig. 1

Optimization of isolated mouse heart mitochondria for bioenergetics assay with XF24 Analyzer (A) Adherence of 1, 2, 3 or 5 µg of mouse heart mitochondria to XF24 v7 cell plate in XF assay. Mouse heart mitochondria were isolated and attached to XF24 microplate as described in Materials and methods. Photomicrograph was taken before the XF assay; (B) mitochondrial coupling assay with 1, 2, 3 or 5 µg of heart mitochondria per well of XF plate as described in Materials and methods. Effect of ADP, oligomycin, FCCP and antimycin A; (C) absolute oxygen levels in the micro-chamber due to mitochondrial coupling using 1, 2, 3 or 5 µg mitochondria per well. (D) Plot of mitochondrial protein against state 2, state 3 and state 3 µ respirations. (E) Mitochondrial electron flow assay using pyruvate and malate as substrates as described in Materials and methods, and the effect of rotenone, succinate, antimycin A and ascorbate/TMPD; (F) plot of mitochondrial protein against the state 3 µ, succinate and Asc/TMPD driven respirations; (G) absolute pH level in the coupling assay. The effect of various amounts of heart mitochondria/well on RCR (state 3/state 40).