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Fig. 1. (A) Differential expression of ß1 integrin epitopes on K562 Cells. Cells were grown in serum-free medium and either stained with JB1A, B3B11 or 12G10, or reacted with only the secondary antibody (BG). The cells were analysed by flow cytometry. The geometric means of the expression values for each category were BG 8. 27, 12G10 29. 97, B3B11 61. 95 and JB1A 72. 27. (B) Distribution of ß1 integrin species identified by B3B11 or 12G10. K562 cells were fixed in 4% paraformaldehyde, stained with the indicated antibodies and visualized with Cy3-conjugated goat anti-mouse immunoglobulin. The cells were stained with a polyclonal antibody 
5 and stained with an Oregon Green goat anti-rabbit immunoglobulin. (C) 12G10-reactive ß1 does not codistribute with B3B11 reactive ß1. Cells were reacted with 12G10 and the bound integrins were cross-linked with Cy3-coupled goat anti-mouse immunoglobulin. The cells were fixed and blocked with mouse immunoglobulin to saturate the combining sites on the anti-mouse immunoglobulin. The cells were then reacted with biotin-labelled B3B11 and the bound antibody was detected with an FITC-labelled Avidin conjugate. 12G10 labelled integrins were clustered; in contrast B3B1ll integrins were diffusely distributed. The overlay of the two images shows that there was little or no colocalization of the integrins detected by these two antibodies.
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