Mouse Anti-ABCG2 Recombinant Antibody (Clone 5D3) (CAT#: PABS-0024)

Figure 1 Alexa Fluor 647 conjugated ABCG2-specific antibody (5D3) was used to detect ABCG2 on the cell surface.

Scale bars represent 10 µm.

Bartos, Z., & Homolya, L. (2021). Identification of specific trafficking defects of naturally occurring variants of the human ABCG2 transporter. Frontiers in Cell and Developmental Biology, 9, 40.

Figure 2 Trafficking of the ABCG2 variants from the ER to the cell surface.

HeLa cells expressing the M71V-ABCG2 variants were labeled with various cellular markers in the absence (0 h) and after the addition of biotin (1-4 h). Representative confocal images show the cellular localization of the ABCG2 variant (green) and the marker (red). Scale bars represent 10 µm.

Bartos, Z., & Homolya, L. (2021). Identification of specific trafficking defects of naturally occurring variants of the human ABCG2 transporter. Frontiers in Cell and Developmental Biology, 9, 40.

Figure 3 Trafficking of the ABCG2 variants from the ER to the cell surface.

HeLa cells expressing the Q141K-ABCG2 variants were labeled with various cellular markers in the absence (0 h) and after the addition of biotin (1-4 h). Representative confocal images show the cellular localization of the ABCG2 variant (green) and the marker (red). Scale bars represent 10 µm.

Bartos, Z., & Homolya, L. (2021). Identification of specific trafficking defects of naturally occurring variants of the human ABCG2 transporter. Frontiers in Cell and Developmental Biology, 9, 40.

Figure 4 The cell surface labeling with 5D3 (G-I) as cellular markers.

K_in represent the time constants for entering the given compartment, respectively. Data represent mean ± SEM of three independent experiments involving 100-120 cells each. Asterisks indicate statistically significant differences (p < 0.05).

Bartos, Z., & Homolya, L. (2021). Identification of specific trafficking defects of naturally occurring variants of the human ABCG2 transporter. Frontiers in Cell and Developmental Biology, 9, 40.

Figure 5 Flow cytometry detection of ABCG2 in the parental PLB (panels A) and the ABCG2-expressing PLB cells (panels C) by the 5D3.

Effect of fixation by PFA and permeabilization by PFA plus Triton X-100. PLB cells were treated with 1% PFA (dashed line) or 4% PFA plus 0.05% Triton X-100 (heavy solid line) before 5D3 labeling. Nontreated PLB cells (native, solid line) were also labeled with one of the monoclonal antibodies or isotype control (IT, dotted line). Fluorescence of phycoerythrin-conjugatedsecondary antibody was analyzed by flow cytometry.

Ozvegy-Laczka, C., Várady, G., Köblös, G., Ujhelly, O., Cervenak, J., Schuetz, J. D., ... & Sarkadi, B. (2005). Function-dependent conformational changes of the ABCG2 multidrug transporter modify its interaction with a monoclonal antibody on the cell surface. Journal of Biological Chemistry, 280(6), 4219-4227.

Figure 6 Flow cytometry detection of ABCG2 in the parental HEK 293 (panels E) and the ABCG2-expressing HEK 293 cells (panels G) by the 5D3.

Cells were incubated with 0.2 g of 5D3 antibody/10⁶ cells without (5D3, solid line) or with the addition of 5 M Ko143 (dashed line) or 2 mM sodium orthovanadate (Vi, heavy solid line). IT means isotype control (dotted line).

Ozvegy-Laczka, C., Várady, G., Köblös, G., Ujhelly, O., Cervenak, J., Schuetz, J. D., ... & Sarkadi, B. (2005). Function-dependent conformational changes of the ABCG2 multidrug transporter modify its interaction with a monoclonal antibody on the cell surface. Journal of Biological Chemistry, 280(6), 4219-4227.

Figure 7 Detection of 5D3 mAb binding to ABCG2 in isolated Sf9 membrane fragments.

Comparison of 5D3 and isotype control (IT) antibody binding to isolated Sf9 cell membranes. Isolated membrane fragments (45 g) from Sf9 cells containing WT ABCG2, ABCG2-K86M, or MDR1 were labeled with 1 μg/ml 5D3 (black columns) or 1 μg/ml mouse IgG2b as isotype control (white columns). Fluorescence was detected in a fluorescence plate reader (Fluoroskan II, Labsystems) at 485 nm (excitation)/590 nm (emission).

Ozvegy-Laczka, C., Várady, G., Köblös, G., Ujhelly, O., Cervenak, J., Schuetz, J. D., ... & Sarkadi, B. (2005). Function-dependent conformational changes of the ABCG2 multidrug transporter modify its interaction with a monoclonal antibody on the cell surface. Journal of Biological Chemistry, 280(6), 4219-4227.