Mitochondrial Markers Antibody Pack

Detection of COX4 in A431, HepG2, NIH-3T3 and recombinant protein by Western Blot.

Western blot shows lysates of A431 human epithelial carcinoma cell line, HepG2 human hepatocellular carcinoma cell line, and NIH-3T3 mouse embryonic fibroblast cell line. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human COX4 Monoclonal Antibody (Catalog # MAB6980) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). For addtional reference, recombinant human COX4-I1 (5 ng/lane) and recombinant human COX4-I2 (2.5 ng/lane) were included. A specific band was detected for COX4 at approximately 18 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

Detection of Cytochrome C in Human HeLa Cell Line by Flow Cytometry.

Human HeLa cell line was stained with Mouse Anti- Cytochrome C Monoclonal Antibody (Catalog # NB100-56503, filled histogram), or Mouse IgG2B isotype control (Catalog # MAB0041, open histogram) followed by APC-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005).Images may not be copied, printed or otherwise disseminated without express written permission of Novus Biologicals a bio-techne brand.

Detection of HSP60 in HeLa cells by Immunocytochemistry/Immunofluorescence.

HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with HSP60 Antibody (NBP1-77397) at 1 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.