Western immunoblot analysis was performed using whole cell lysates derived from HGSOC cell lines as previously described with minor alterations [6 (link)]. Forty μg of protein lysate/cell line was analyzed. Equal loading was verified by incubating the membranes with anti-GAPDH antibody. Primary antibodies used include monoclonal mouse anti-GAPDH (GeneTex, Irvine, CA, USA), Cat #GTX627408, polyclonal rabbit anti-SUSD2 (C-term. fragment; Prestige Antibodies Sigma-Aldrich Corp.), monoclonal mouse anti-SUSD2 (C-term. fragment; Bio-Techne Corp.) Cat MAB9056, polyclonal rabbit anti-SUSD2 (N-term. fragment; AbCam technologies, San Francisco, CA, USA) Cat ab182147.
For immunoblotting performed in the characterization of SUSD2 expression in stable HGSOC cells, detection was achieved with IRDye-conjugated secondary antibodies (LI-COR Biosciences) and imaged using the infrared Odyssey system; 800CW donkey anti-mouse antibody, and a 680RD donkey anti-rabbit antibody. Experiments were performed using three biological replicates for each cell line.
For immunoblotting performed in the characterization of SUSD2 expression in stable HGSOC cells, detection was achieved with IRDye-conjugated secondary antibodies (LI-COR Biosciences) and imaged using the infrared Odyssey system; 800CW donkey anti-mouse antibody, and a 680RD donkey anti-rabbit antibody. Experiments were performed using three biological replicates for each cell line.
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