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After incubation with purified GST-PLSCR1 or GST, binding was revealed with an anti-GST antibody.
Primary Ab binding was revealed with biotinylated mouse anti-rabbit Ab (1∶250) for 30 min at RT and then with StreptABcomplex and Alkaline phosphatase (Dako, Glostrup, Denmark) for 30 min at RT, according to manufacturer's instruction.
TARP γ-2 binding was revealed with an anti-γ-2 antibody (Supplemental Experimental Procedures).
Primary antibody binding was revealed with FITC-labelled goat-anti-mouse or anti-rabbit antibodies (Dianova GmbH).
After incubating with dilutions of the patient's serum samples, binding was revealed with an anti-idiotype antibody directed against the humanised anti-CEACAM5 portion of TF2 (Sharkey et al, 2010).
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Antibody binding was revealed by incubation with anti-mouse (Sigma) or anti-rabbit (Santa Cruz Biotechnology) IgG peroxidase conjugate secondary antibodies.
Antibody binding was revealed by incubation with horseradish peroxidase-conjugated secondary antibodies (GE Biosciences) and the ECL Plus immunoblotting detection system (GE Biosciences).
Antibody binding was revealed by incubation with horseradish peroxidase-conjugated secondary antibodies followed by ECL Plus immuno-blotting detection system (Amersham Biosciences).
ScFv binding was revealed by 60 min. incubation with NitroBlue Tetrazolium/Bromo-chloro Indoyl Phosphate (NBT/BCIP) substrate.
Antibody binding was revealed using 3,3´-diaminobenzidine, and slides were counterstained with hematoxylin.
Wells were probed with a TF2-horseradish peroxidase conjugate, and binding was revealed using o-phenylenediamine dihydrochlorides.
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