Characterization of anti-fls485 antibodies. Detection of chimeric EGFP-fls485158 protein in transfected 3T3 (A: Western blot) or CaCo2 cells (B-H: immunofluorescence) with different anti-fls485 antibodies. (A) Western blots of EGFP-fls485158 expressed in 3T3 cells incubated with antibodies directed against fls485158 clone/subclone #7 (left) or clone/subclone #10 (middle). Endogenous fls485 expression by 3T3 cells is detectable as a signal about 55 kDa when incubated with the anti-fls485 clones. One additional signal about 35 kDa is exclusively found when clone/subclone #10 is used. EGFP-fls485158 expression in 3T3 transfectants was additionally visualized with anti-EGFP antibodies as control (right). (B-D) Immunofluorescence of EGFP-fls485158 protein in transfected CaCo2 cells incubated with clone/subclone #7, secondary antibody Cy3-labeled (B), anti-EGFP, secondary antibody FITC-labeled (C), and overlay (D). (E-G) Immunofluorescence of EGFP-fls485158 protein in transfected CaCo2 cells incubated with clone/subclone #10, secondary antibody Cy3-labeled (E), anti-EGFP, secondary antibody FITC-labeled (F), and overlay (G). (H) Negative control; EGFP-fls485158 transfected CaCo2 cells after incubation with all secondary antibodies.