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. 2023 Oct 31;24(21):15847. doi: 10.3390/ijms242115847

Figure 1.

Figure 1

Analysis of several commercial anti-ABCB5 antibodies to analyze ABCB5 expression in MelJuSo cells. (A) Western blotting detection of human ABCB5β in membrane vesicles of high-five insect cells infected or not with ABCB5β-containing baculoviruses. Total membranes (1 µg) were resolved by SDS-PAGE prior to transfer on a PVDF membrane. Three different commercial anti-ABCB5 antibodies were tested: rabbit polyclonal anti-ABCB5 (Rockland Immunochemicals, Limerick, PA, USA, 600-401-A775), rabbit anti-ABCB5 (Abcam, Cambridge, UK, ab8010889), and rabbit anti-ABCB5 (Atlas antibodies, Lund, Sweden, HPA026975). (B) RT-qPCR analysis of ABCB5 mRNA expression in MelJuSo cells after transfection with a scrambled siRNA pool (siCtrl) or a pool of 4 siRNAs directed against ABCB5 (siABCB5) over 24 h, 48 h, 72 h, or 96 h. Primers recognize a sequence located in the beta isoform. mRNA expression levels normalized to GAPDH and relative to siCtrl conditions are shown on the graph. (C) Western blotting detection of endogenous ABCB5 protein expression in MelJuSo cells transfected with siCtrl or siABCB5 over 24 h, 48 h, 72 h, and 96 h, using the 3 different commercial anti-ABCB5 antibodies anti-ABCB5 (Rockland Immunochemicals, Limerick, PA, USA, 600-401-A775), rabbit anti-ABCB5 (Abcam, Cambridge, UK, ab8010889), and rabbit anti-ABCB5 (Atlas antibodies, Lund, Sweden, HPA026975).