Samples were separated by SDS-PAGE using pre-casted 4–20% acrylamide gels (Bio-Rad, Hercules, CA, USA) and proteins transferred to nitrocellulose membranes. Membranes were blocked with 5% milk, incubated with primary antibody overnight followed with HRP-conjugated secondary antibody. Signal was developed using ECL and exposure to X-ray film. Antibodies used in this study were obtained from Abcam/Mitosciences (NDUFA9, NDUFB8, UQCRC1, UQCRC2, UQCRFS1, Cox1, Cox5b, SDHA and ATP5A); from Sigma (NDUFV1 and Actin); from Research Diagnostics Inc. (SOD1); from Antibody Verify (SOD2); from BD Bioscience (Tim23); and from Abgent (GPx1). All antibodies were used at a 1:1000 dilution in diluted blocking solution (0.25% w/w milk in PBST). Blots were quantified by densitometry were performed using ImageJ 1.49n software (National Institute of Health, Bethesda, MD, USA) and Tim23 or actin signals were used for normalization.
Cox5b
Manufactured by Merck Group
Sourced in Germany
Cox5b is a laboratory equipment product from Merck Group. It is a component of the mitochondrial electron transport chain, specifically the cytochrome c oxidase complex. Cox5b plays a core function in cellular respiration and energy production.
Automatically generated - may contain errors
2 protocols using cox5b
1
Western Blot Analysis of Mitochondrial Proteins
2
Immunohistochemical Profiling of Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Immunostaining was performed on 4-mm-thick sections of formalin-fixed, paraffin-embedded specimen after deparaffinization and microwave-based antigen retrieval as previously described. 16 The IView DAB Detection Kit (Roche, Basel, Switzerland) was used on a Ventana BenchMark platform (Roche) for the following antibodies: CD4 (Clone SP35; Roche), CD8 (Clone SP57; Roche), forkhead box P3 (Clone 236A/E7; Thermo Fisher Scientific, Bremen, Germany), and cytochrome c oxidase subunit 5B (COX5B) (Clone 1E8; Sigma-Aldrich, Taufkirchen, Germany). Immunostaining for glucose transporter 1 (GLUT1) (Clone SPM498; LabVision, Fremont, CA) was performed manually with an appropriate horseradish peroxidaseeconjugated secondary antibody.
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