Fig.1: FACS analysis of BOSC23 cells using HT-12D8 Cat.# GM-0516. BOSC23 cells were transiently transfected with an expression vector encoding either CEACAM20 (red curve) or an irrelevant protein (control transfectant). Binding of HT-12D8 was detected with a PE- conjugated secondary antibody. A positive signal was obtained only with CEACAM20 transfected cells.
Fig.2: Spectral Confocal Microscopy of CHO cells using HT-12D8 Cat.# GM-0516. CHO cells were transiently transfected with an expression vector encoding CEACAM20. Binding of HT-12D8 was visualized with a FITC-conjugated secondary antibody (green). Actin filaments are labeled with Alexa Fluor-555 Phalloidin (red). Cell nuclei are stained with DAPI (blue).
Fig3: BOSC23 cells were transiently transfected with expression vectors containing either the cDNA of CEACAM1, CEACAM3-8 or CEACAM19-21. Expression of the constructs was tested with monoclonal antibodies known to recognize the corresponding proteins (CEACAM1,3,4,5 and 6: D14HD11; CEACAM7: BAC2; CEACAM8: Tet2; CEACAM19,21: ?-myc; CEACAM20: ?-flag; green curves). An irrelevant monoclonal antibody served as a negative control (black curves). For specificity testing, protein G-purified HT-12D8 was tested on all CEACAM transfectants. A positive signal was obtained only with CEACAM20 transfected cells (red curve).
SDS-PAGE analysis of HT-12D8 The antibody was purified by protein G affinity chromatography from cell culture supernatants and verified by SDS-Page (Fig.4). Fig.4: SDS-PAGE analysis of purified HT-12D8 monoclonal antibody. Lane 1: molecular weight marker, Lane 2: 2 ?g of purified HT-12D8 antibody. Proteins were separated by SDS-PAGE and stained with RAPID StainTM Reagent.
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