Protein Extraction and Western Blot Analysis

Using a protein extraction kit (Applygen Technologies), total protein was extracted and the concentration of protein was determined by a BCA protein assay kit (Applygen Technologies). Western blot analysis was performed routinely (10 (link)), with primary antibodies against NDUFA10 (Santa Cruz, Santa Cruz, Calif), adenosine triphosphate synthase δ-subunit (ATP 5D) (Abcam), Sirt1, Sirt3, SDHA, B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), cleaved caspase-3, and glyceraldehyde-3-phosphatedehydrogenase (GAPDH) (Cell Signaling Technology, Beverly, Mass). Enhanced chemiluminescence detection kit (Applygen Technologies, Beijing, China) was used to detect the bands. Band intensity was expressed as mean area density using Quantity One image analyzer software (Bio-Rad, Richmond, Calif) for quantification.

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Huang D.D., Wei X.H., Mu H.N., Pan C.S., Li Q., Hu B.H., Chang X., Yan L., Fan J.Y., Liu Y.Y., Luo J.Y, & Han J.Y. (2019). Total Salvianolic Acid Injection Prevents Ischemia/Reperfusion-Induced Myocardial Injury Via Antioxidant Mechanism Involving Mitochondrial Respiratory Chain Through the Upregulation of Sirtuin1 and Sirtuin3. Shock (Augusta, Ga.), 51(6), 745-756.

Publication 2019

protein extraction kit BCA protein assay kit glyceraldehyde-3-phosphatedehydrogenase (GAPDH) Enhanced chemiluminescence detection kit Quantity One image analyzer software

Adenosine triphosphate Antibodies Assay B cell lymphoma Caspase 3 Chemiluminescence Gapdh Glyceraldehyde Protein Protein a Sirt1 Sirt3 Subunit Synthase Western blot analysis

Corresponding Organization : Shanghai Innovative Research Center of Traditional Chinese Medicine

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Variable analysis

independent variables

Protein extraction kit (Applygen Technologies)
Primary antibodies against NDUFA10 (Santa Cruz, Santa Cruz, Calif), adenosine triphosphate synthase δ-subunit (ATP 5D) (Abcam), Sirt1, Sirt3, SDHA, B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), cleaved caspase-3, and glyceraldehyde-3-phosphatedehydrogenase (GAPDH) (Cell Signaling Technology, Beverly, Mass)

dependent variables

Total protein concentration determined by a BCA protein assay kit (Applygen Technologies)
Protein band intensity measured using Quantity One image analyzer software (Bio-Rad, Richmond, Calif)

control variables

Western blot analysis performed routinely (10 (link))
Enhanced chemiluminescence detection kit (Applygen Technologies, Beijing, China) used to detect the bands

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