Western Blot Analysis of Tissue Samples

Tissue samples were processed for Western blotting using our standard method, as described.7 Briefly, tissue samples (brain cortex, spinal cord, and kidney) were dissected on ice and snap frozen in liquid nitrogen as quickly as possible to prevent deconjugation of SUMOylated and ubiquitinated proteins. Tissue samples were homogenized by sonication using lysis buffer supplemented with 2% SDS. Quantification of signal intensities was performed using ImageJ (NIH, Bethesda, MD). The primary antibodies are listed in Table 2.

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Shen Y., Yan B., Zhao Q., Wang Z., Wu J., Ren J., Wang W., Yu S., Sheng H., Crowley S.D., Ding F., Paschen W, & Yang W. (2018). Aging Is Associated With Impaired Activation of Protein Homeostasis‐Related Pathways After Cardiac Arrest in Mice. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 7(17), e009634.

Publication 2018

Antibodies Brain Buffer Cortex Frozen Kidney Nitrogen Spinal cord Tissue Ubiquitinated proteins

Corresponding Organization :

Other organizations : Duke Medical Center, Nantong University, Second Affiliated Hospital of Xi'an Jiaotong University, Duke University Hospital, Tianjin Medical University General Hospital, Duke University, Durham VA Medical Center, Nanfang Hospital, Southern Medical University

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Variable analysis

independent variables

Tissue sample type (brain cortex, spinal cord, and kidney)

dependent variables

Signal intensities of SUMOylated and ubiquitinated proteins

control variables

Tissue processing (dissection on ice, snap freezing in liquid nitrogen)
Homogenization method (sonication using lysis buffer with 2% SDS)
Quantification method (ImageJ)

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