Fig 1: Western blot of recombinant Hsp70 protein and tumor cell lysates, flow cytometric analysis of membrane-bound Hsp70 on viable tumor cells, and dot blot analysis using BSA, recombinant Hsp27, Hsp60, Hsp70, Hsc70 as target proteins. (a) Hsp70 protein and lysates of tumor cells were subjected to an SDS gel and blots were stained with cmHsp70.1, cmHsp70.2, ctrl Hsp70A and ctrl Hsp70B antibodies. Representative immunoblots of recombinant Hsp70 (upper, 72 kDa) and tumor cell lysates (lower) representing Hsp70 (72 kDa), Hsc70 (73 kDa) and β-actin (40 kDa) as a loading control are shown. Molecular weight markers are indicating 70 kDa and 40 kDa. Detailed information about Western blot can be found at Supplementary Materials. (b) Detection of membrane-bound Hsp70 on A549, HCT116 cell lines by flow cytometry using the cmHsp70.1, cmHsp70.2, ctrl Hsp70A and ctrl Hsp70B antibodies (white histograms). Staining with the respective isotype-matched control antibodies is represented by grey histograms. As a control, PBL of healthy donors were stained with cmHsp70.1 and cmHsp70.2 mAb. The numbers in the histograms indicate the proportion of Hsp70 positively stained cells. (c) Dot blot analysis of BSA, recombinant Hsp27, Hsp60, Hsp70 and Hsc70 (100 ng, 10 ng, 1 ng, 0.1 ng) using cmHsp70.1, cmHsp70.2 and ctrl Hsp70B antibodies.