Hrp conjugated anti goat igg antibody

Immunoblot analyses were carried out as previously described [15 (link)]. We used rabbit polyclonal anti-ALAD (Abcam, Cambridge, UK), mouse monoclonal anti-Actin (MP Biomedicals, Santa Ana, California, USA), mouse monoclonal anti-COX4I1 (Santa Cruz Biotechnology, Dallas, Texas, USA), goat polyclonal anti-HIF-1α (R&D systems, Minneapolis, USA), rabbit polyclonal anti-HSP90 (Santa Cruz Biotechnology, Dallas, Texas, USA), and mouse monoclonal anti-PPOX (Abcam, Cambridge, UK) as the primary antibodies, depending on the purpose of the immunoblot analysis. For the second antibody, we used horseradish peroxidase (HRP)-conjugated anti-mouse IgG antibody, HRP-conjugated anti-rabbit IgG antibody (Cell Signaling Technology, Beverly, Massachusetts, USA), and HRP-conjugated anti-goat IgG antibody (Santa Cruz Biotechnology, Dallas, Texas, USA) at 1:3000 dilution.

Cells were lysed in the SDS-sample buffer, separated by SDS-PAGE and transferred to a PVDF membrane. Immunoblotting was performed with a goat anti-HRP antibody (1∶5000; Jackson ImmunoResearch). HRP-conjugated anti-goat IgG antibody (Santa Cruz Biotechnology) was diluted 10,000-fold and used as a secondary antibody.

Unless otherwise indicated, all of the chemicals were purchased from Sigma (St. Louis, MO, USA). All of antibodies were stored at 4 °C until use: rabbit anti-S100 antibody (Dako Cytomation, Denmark), mouse anti-COX IV antibody (Abcam, Cambridge, UK), goat anti-ABCB5 antibody (Abnova, Taipei, Taiwan), Alexa-488-conjugated and Alexa-546-conjugated secondary antibodies (Molecular Probe, Eugene, OR, USA), rabbit anti-CD133 antibody, mouse anti-Ki-67 antibody, horseradish peroxidase-(HRP)-conjugated anti-mouse IgG antibody, HRP-conjugated anti-rabbit IgG antibody and HRP-conjugated anti-goat IgG antibody (Santa Cruz, CA, USA).