Quantifying SIRT1 Protein Levels

Frozen placental tissue samples were homogenized in liquid nitrogen and lysed in cell lysis buffer (Beyotime Biotechnology, Shanghai, China). Protein content was measured with the BCA kit (Beyotime Biotechnology, Shanghai, China). The Western blot analysis steps were conducted according to previously reported methods [38 (link)]. Primary antibodies against SIRT1(1:1000, 9475S, CST, Danvers, MA, USA) and GAPDH (1:1000, abs132004, Absin, Shanghai, China) were used in this study. Blots were analyzed with ImageJ software (NIH, Bethesda, MD, USA).

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Peng X., Cai X., Li J., Huang Y., Liu H., He J., Fang Z., Feng B., Tang J., Lin Y., Jiang X., Hu L., Xu S., Zhuo Y., Che L, & Wu D. (2021). Effects of Melatonin Supplementation during Pregnancy on Reproductive Performance, Maternal–Placental–Fetal Redox Status, and Placental Mitochondrial Function in a Sow Model. Antioxidants, 10(12), 1867.

Publication 2021

cell lysis buffer BCA kit abs132004

Antibodies Buffer Cell Frozen Gapdh Nitrogen Placental Protein Sirt1 Tissue Western blot analysis

Corresponding Organization : Sichuan Agricultural University

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Variable analysis

independent variables

Frozen placental tissue samples homogenized in liquid nitrogen

dependent variables

SIRT1 protein expression
GAPDH protein expression

control variables

Cell lysis buffer (Beyotime Biotechnology, Shanghai, China)
Protein content measurement with BCA kit (Beyotime Biotechnology, Shanghai, China)
Western blot analysis steps conducted according to previously reported methods [38]

controls

Positive control: Not specified
Negative control: Not specified

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