Journal: PLoS ONE
Article Title: Validation of commercial Mas receptor antibodies for utilization in Western Blotting, immunofluorescence and immunohistochemistry studies
Figure Lengend Snippet: Immunofluorescence studies in HEK293T cells overexpressing c-myc tagged MasR using MasR antibodies. Images of fluorescence signal corresponding to secondary cyanine (Cy3)-conjugated antibody (A), nuclear Hoechst 33258 staining (B), merged images (C) and bright field (D) are shown. In cells transfected with the pcDNA 3.1/c-myc-MasR construct, antibodies NLS-1531 and sc-54682 generated a similar staining pattern to that generated with the anti c-myc antibody. No staining was observed with any of these antibodies in cells transfected with the empty vector pcDNA 3.1. Antibody sc-135063 was capable of staining the plasma membrane of cells overexpressing the MasR but also generated widespread signals in cells transfected with the empty vector. For antibody AAR-013 there was no staining associated with the cell membrane. Intense immunocytochemical staining of identical distribution and intensity was revealed in cells transfected with the empty vector and those transfected with the c-myc-MasR construct. Images are representative of 3 independent experiments.
Article Snippet: Two antibodies (sc-135063 and sc-54682) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA); AAR-013 was from Alomone Labs (Jerusalem, Israel) and NLS1-1531 was from Novus Biologicals (Littleton, CO, USA).
Techniques: Immunofluorescence, Fluorescence, Staining, Transfection, Construct, Generated, Plasmid Preparation