Immunoblotting of Renal Cortex Proteins

Proteins of interest in renal cortex homogenates were detected by immunoblotting as described previously [36 (link)]. Primary antibodies were against: AS-β, neutrophil gelatinase-associated lipocalin (NGAL), Tfam (GenWay Biotech, San Diego, CA), ED-1 (Serotek, Raleigh, NC), myeloperoxidase (MPO; DAKO, Carpinteria, CA), cleaved caspase-3 (Cell Signaling Technology, Danvers, MA), dynamin-related GTPase protein Drp-1, fissin-1 (Fis-1), mitofusin-1 (Mfn-1), ND3, PGC-1α, PTEN-induced putative kinase 1 (PINK-1) (Santa Cruz Biotech., Santa Cruz, CA) and microtubule-associated protein 1A/1B-light chain 3 (LC3, MBL International, Des Plainers, IL) at concentrations of 1:100 to 1000, and actin (ICN, Costa Mesa, CA) at a concentration of 1:3000. Detection was achieved by chemiluminescence (Pierce Biotechnology, Rockford, IL).

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Liu Q., Krishnasamy Y., Rehman H., Lemasters J.J., Schnellmann R.G, & Zhong Z. (2015). Disrupted Renal Mitochondrial Homeostasis after Liver Transplantation in Rats. PLoS ONE, 10(10), e0140906.

Publication 2015

myeloperoxidase (MPO cleaved caspase-3 PGC-1α

Actin Antibodies Caspase 3 Chemiluminescence Costa Dynamin related protein 1 Gtpase Kinase Light Microtubule associated protein 3 Myeloperoxidase Neutrophil gelatinase associated lipocalin Pgc 1α Proteins Pten Renal cortex Tfam

Corresponding Organization : Ralph H. Johnson VA Medical Center

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Variable analysis

independent variables

Primary antibodies used for immunoblotting

dependent variables

Detection of proteins of interest in renal cortex homogenates by immunoblotting

control variables

Actin (used as a loading control)

controls

Positive control: None explicitly mentioned
Negative control: None explicitly mentioned

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