Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

2.13. Measurement of mitochondrial oxygen consumption rate using a Seahorse analyzer

CS Chi-Chin Sun
SL Shao-Yun Lee
CK Cheng-Heng Kao
LC Li-Hsien Chen
ZS Zhao-Qing Shen
CL Chia-Hui Lai
TT Tsai-Yu Tzeng
JP Jong-Hwei Su Pang
WC Wen-Tai Chiu
TT Ting-Fen Tsai
request Request a Protocol
ask Ask a question
Favorite

An XFe24 analyzer (Seahorse Bioscience, MA) was applied to measure the oxygen consumption rate (OCR) of mitochondria. The HCEC cells were cultured in a fibronectin-coated (0.01 mg/mL fibronectin, 0.03 mg/mL bovine collagen type I and 0.01 mg/mL bovine serum albumin) XF24 V7 plate at 3×104 cells/well. The HCEC cells were kept overnight in HCEC complete medium and then replaced by fresh HCEC complete medium (pH=7.4) at 1 hr before measurement. The OCR was measured before and after added the indicated chemicals (1 μM oligomycin A, 2 μM FCCP and 1 μM rotenone with 1 μM antimycin A) at 37°C. The results are presented in pmol/min/μg protein.

Copyright and License information:  ©2021 The Authors
This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

post Post a Question
0 Q&A