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Molecules that bind B7.1 can be isolated from plasma using CTLA-4 specific antibodies . CTLA-4 positive plasma, from subjects 19 (A), 22 (B), and 44 (C), were passed through a column linked to the CTLA-4 specific antibody AS32. Fractions, containing material that eluted from the column at low pH, were bound to 96 well plates and tested for interaction with various biotinylated proteins by ELISA: B7.1-Ig (gray bars), Muc-Ig (black bars), and the AS33 antibody (circles).

Journal: BMC Immunology

Article Title: Biochemical analysis of CTLA-4 immunoreactive material from human blood

doi: 10.1186/1471-2172-10-51

Figure Lengend Snippet: Molecules that bind B7.1 can be isolated from plasma using CTLA-4 specific antibodies . CTLA-4 positive plasma, from subjects 19 (A), 22 (B), and 44 (C), were passed through a column linked to the CTLA-4 specific antibody AS32. Fractions, containing material that eluted from the column at low pH, were bound to 96 well plates and tested for interaction with various biotinylated proteins by ELISA: B7.1-Ig (gray bars), Muc-Ig (black bars), and the AS33 antibody (circles).

Article Snippet: B7.1-Ig (CD-80) and B7.2-Ig (CD86) fusion proteins (R&D Systems, Minneapolis, MN) were biotinylated with the use of a commercially available kit (Pierce Immunochemical, Rockford, IL).

Techniques: Isolation, Enzyme-linked Immunosorbent Assay

Material that binds B7.1 and B7.2 can only be isolated from blood that contains molecules recognized by CTLA-4 specific antibodies . Plasma from subjects 17 (CTLA-4 negative) and 19 (CTLA-4 positive) were passed over a column linked to the CTLA-4 specific antibody AS32. Material that eluted from the column under stringent wash conditions was collected in fractions, and was used to coat the wells of a 96 well plate. In panel (A) these wells were probed with the CTLA-4 specific antibody AS33. Panel (B) shows the binding of B7.1-Ig, B7.2-Ig, and Muc-Ig to the coated wells.

Journal: BMC Immunology

Article Title: Biochemical analysis of CTLA-4 immunoreactive material from human blood

doi: 10.1186/1471-2172-10-51

Figure Lengend Snippet: Material that binds B7.1 and B7.2 can only be isolated from blood that contains molecules recognized by CTLA-4 specific antibodies . Plasma from subjects 17 (CTLA-4 negative) and 19 (CTLA-4 positive) were passed over a column linked to the CTLA-4 specific antibody AS32. Material that eluted from the column under stringent wash conditions was collected in fractions, and was used to coat the wells of a 96 well plate. In panel (A) these wells were probed with the CTLA-4 specific antibody AS33. Panel (B) shows the binding of B7.1-Ig, B7.2-Ig, and Muc-Ig to the coated wells.

Article Snippet: B7.1-Ig (CD-80) and B7.2-Ig (CD86) fusion proteins (R&D Systems, Minneapolis, MN) were biotinylated with the use of a commercially available kit (Pierce Immunochemical, Rockford, IL).

Techniques: Isolation, Binding Assay