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Image Search Results
Journal: Journal of Biological Chemistry
Article Title: Mapping of a Conformational Epitope on Plasminogen Activator Inhibitor-1 by Random Mutagenesis
doi: 10.1074/jbc.m208420200
Figure Lengend Snippet: FIG. 1. Screen of 33B8 binding deficient PAI-1 mutants. Panels A–C, filter series showing EX(lox) plaques. Panel A, plaques immunore- active with murine monoclonal 33B8; panel B, plaques reactive with rabbit anti-PAI-1 polyclonal antibodies; panel C, a merged image of A and B. The same filter is shown in each case. The dark spots indicate PAI-1antibody complexes, and the circles indicate examples of PAI-1 mutants that are specifically disrupted in 33B8 binding. Panels D and E show immunoblots of cell-free extracts of wild-type and parental PAI-1 mutants selected from the screen. Panel D was probed with 33B8 and panel E with anti-PAI-1 polyclonal antibodies. Panel F shows the inhibitory activity toward high molecular weight uPA of wild-type and each of the PAI-1 parental mutants in cell-free extracts. The inhibitory activity is expressed as the percentage of active PAI-1 protein relative to the total soluble protein of the cell extracts.
Article Snippet: Materials—Murine monoclonal antibodies MA-33B8 (33B8) and MA31C9 (31C9) as well as rabbit polyclonal antibodies directed against human PAI-1 were purchased from Molecular Innovations (Southfield, MI), horseradish peroxidase-conjugated goat anti-mouse and anti-rabbit antiserum were from Bio-Rad. tPA was from Genentech (South San Francisco, CA), and
Techniques: Binding Assay, Western Blot, Activity Assay, High Molecular Weight
Journal: Journal of Biological Chemistry
Article Title: Mapping of a Conformational Epitope on Plasminogen Activator Inhibitor-1 by Random Mutagenesis
doi: 10.1074/jbc.m208420200
Figure Lengend Snippet: FIG. 4. Analysis of site-directed PAI-1 mutants. Panels A and B show immunoblots of cell-free extracts of wild-type and site-directed PAI-1 mutants. Panel A was probed with 33B8 and panel B with anti-PAI-1 polyclonal antibodies. Panel C shows the inhibitory activity toward high molecular weight uPA of wild-type and each of the PAI-1 site-directed mutants in cell-free extracts. The inhibitory activity is expressed as the percentage of functionally active PAI-1 relative to the total soluble protein of the cell extracts.
Article Snippet: Materials—Murine monoclonal antibodies MA-33B8 (33B8) and MA31C9 (31C9) as well as rabbit polyclonal antibodies directed against human PAI-1 were purchased from Molecular Innovations (Southfield, MI), horseradish peroxidase-conjugated goat anti-mouse and anti-rabbit antiserum were from Bio-Rad. tPA was from Genentech (South San Francisco, CA), and
Techniques: Western Blot, Activity Assay, High Molecular Weight