Mitochondria Isolation from HEK293 Cells

Mitochondria were isolated from HEK293 cells as previously described [18 (link)]. Briefly, 60 cm plates of HEK293 cells were collected by adding 5 mL of homogenization buffer (300 mmol/L sucrose, 10 mmol/L HEPES-KOH, 1 mmol/L EGTA-KOH, pH 7.4) with 1 mg Subtilisin A protease from Bacillus licheniformis (Sigma-Aldrich). A detailed protocol for mitochondria isolation can be found in our previous published manuscript [18 (link)].

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Namwanje M., Bisunke B., Rousselle T.V., Lamanilao G.G., Sunder V.S., Patterson E.C., Kuscu C., Kuscu C., Maluf D., Kiran M., Mas V., Eason J.D, & Bajwa A. (2021). Rapamycin Alternatively Modifies Mitochondrial Dynamics in Dendritic Cells to Reduce Kidney Ischemic Reperfusion Injury. International Journal of Molecular Sciences, 22(10), 5386.

Publication 2021

Subtilisin A protease from Bacillus licheniformis

Bacillus licheniformis Buffer Egta Hek293 cells Hepes Isolation Mitochondria Protease Subtilisin a Sucrose

Corresponding Organization : University of Tennessee Health Science Center

Other organizations : University of Maryland, Baltimore, University of Hyderabad, University of Maryland Medical Center

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Variable analysis

independent variables

Addition of 1 mg Subtilisin A protease from Bacillus licheniformis

dependent variables

Mitochondria isolated from HEK293 cells

control variables

Homogenization buffer (300 mmol/L sucrose, 10 mmol/L HEPES-KOH, 1 mmol/L EGTA-KOH, pH 7.4)
HEK293 cells

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

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