Immunofluorescence Staining of Frozen Kidney Sections

Immunofluorescence staining was performed with routine protocol [72 (link)]. Frozen kidney sections (3 μm) were fixed with 4% paraformaldehyde. All primary antibodies used were as follows: anti-TOMM20 (ab186735; Abcam), anti-Sirtuin 6 (PB0375; Boster), anti-PGC-1α (ab54481; Abcam). Then the slides were stained with Cy3-or Cy2-conjugated secondary antibodies (Jackson Immuno-Research Laboratories, West Grove, PA, United States). Nuclei were treated with DAPI (Sigma-Aldrich) under the manufacturer’s specifications.

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Miao J., Huang J., Liang Y., Zhang Y., Li J., Meng P., Shen W., Li X., Wu Q., Wang X., Niu H., Tang Y., Zhou S, & Zhou L. (2023). Sirtuin 6 is a key contributor to gender differences in acute kidney injury. Cell Death Discovery, 9, 134.

Publication 2023

ab186735 PB0375 ab54481 Cy3-or Cy2-conjugated secondary antibodies DAPI

Antibodies Dapi Frozen sections Immunofluorescence Kidney Nuclei Paraformaldehyde Pgc 1α Sirtuin 6 Tomm20

Corresponding Organization : Third Affiliated Hospital of Southern Medical University

Other organizations : Nanfang Hospital

Top 5 similar protocols

Variable analysis

independent variables

Immunofluorescence staining protocol

dependent variables

Localization and expression of TOMM20, Sirtuin 6, and PGC-1α proteins

control variables

Frozen kidney sections (3 μm)
4% paraformaldehyde fixation
Primary antibodies: anti-TOMM20, anti-Sirtuin 6, anti-PGC-1α
Cy3- or Cy2-conjugated secondary antibodies
DAPI nuclear staining

controls

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

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