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. 2012 Aug;31(4):267–271. doi: 10.1089/hyb.2012.0008

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Copyright 2012, Mary Ann Liebert, Inc.

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FIG. 1. — Preparation of mouse B7-H3 transfectants and characterization of B7-H3 MAb. (A) CHO/B7-H3 and 293/B7-H3 transfectants were selected and identified by flow cytometry using PE-labeled M3.2D7 (red histograms) against PE-conjugated rat IgG2b (open histograms). (B) Reactivity of the MAbs with the transfectants CHO/mock, CHO/B7-H3, CHO/TLT2, 293/mock, 293/B7-H3, and 293/B7-H4. Cells were stained with MAbs 18F9 and M3.2D7 (red histograms) or negative control rat IgG2b (open histograms). (C) Purified B7-H3-Ig was separated on 10% SDS-polyacrylamide gels, transferred to NC membrane, and stained with PBS, rat IgG2b, M3.2D7, and 18F9. (D) The Ig isotype of the MAb was identified with carboxyl blue-based bead assay. Negative control and 18F9 staining were shown in figure at left (without hybridoma supernatant) and figure at right (with hybridoma supernatant), respectively. The figure is representative of four independent experiments.