Journal:

Article Title: CD4-Induced T-20 Binding to Human Immunodeficiency Virus Type 1 gp120 Blocks Interaction with the CXCR4 Coreceptor

doi: 10.1128/JVI.78.10.5448-5457.2004

Figure Lengend Snippet: Effects of T-20 on antibody binding to HXBc2 gp120. (A and B) HXBc2 gp120 glycoproteins in the supernatant of transiently expressing 293T cells were captured on ELISA plates by a sheep antibody (D7324) against the carboxyl-terminal 15 amino acids of gp120. (A) Bound gp120 glycoproteins were incubated with various concentrations of T-20 peptide and the indicated concentrations of CD4-Ig at room temperature for 1 h. The bound CD4-Ig was detected with an HRP-conjugated anti-Ig secondary antibody and TMB substrates. (B) Bound gp120 glycoproteins were incubated with various concentrations of T-20 peptide in the absence (bottom right) or presence (+sCD4; 2 μg/ml) of sCD4 at room temperature for 1 h, followed by incubation with the indicated concentrations of antibodies at room temperature for another hour. The antibodies bound to gp120 were detected with an HRP-conjugated anti-Ig secondary antibody and TMB substrates. The means and standard deviations of the optical densities at 450 nm (O.D.450) are shown. (C) Radiolabeled HXBc2 gp120 was incubated with sCD4, the indicated amounts of antibodies (Ab), and biotinylated DP178-Ig. Avidin-Sepharose was used for precipitation, and the gp120 bound to the beads was analyzed by SDS-PAGE and autoradiography.

Article Snippet: This soluble DP178-Ig protein was expressed by transfecting the plasmid into 293T cells with GenePorter II reagent (GTS Inc., San Diego, Calif.), and purified from the supernatant by a protein A-Sepharose column.

Techniques: Binding Assay, Expressing, Enzyme-linked Immunosorbent Assay, Incubation, Avidin-Biotin Assay, SDS Page, Autoradiography

Journal:

Article Title: CD4-Induced T-20 Binding to Human Immunodeficiency Virus Type 1 gp120 Blocks Interaction with the CXCR4 Coreceptor

doi: 10.1128/JVI.78.10.5448-5457.2004

Figure Lengend Snippet: Sequence requirements on the T-20/DP178 peptide for gp120 recognition. (A) Radiolabeled YU2 sgp130(−/GCN4) glycoproteins were incubated with PS from HIV-1-infected individuals, DP178-Ig, or C34-Ig and protein A-Sepharose. In parallel, radiolabeled HXBc2 gp120 glycoproteins, in the absence (−) or presence (+) of sCD4, were incubated with PS, DP178-Ig, or C34-Ig. The precipitated proteins were analyzed by SDS-PAGE and autoradiography. (B) Radiolabeled HXBc2 gp120 was incubated with DP178-Ig and protein A-Sepharose in the presence of the indicated concentrations of T-20 or scrT-20. The precipitated gp120 glycoprotein was analyzed by SDS-PAGE and autoradiography.

Article Snippet: This soluble DP178-Ig protein was expressed by transfecting the plasmid into 293T cells with GenePorter II reagent (GTS Inc., San Diego, Calif.), and purified from the supernatant by a protein A-Sepharose column.

Techniques: Sequencing, Incubation, Infection, SDS Page, Autoradiography