DNA for HIV gag-p24 (aa 133–363 derived from HIV isolate BH10) was cloned in frame into the COOH terminus of the heavy chains of α–mouse-DEC205 as described previously (19 (link)). The fusion α–CD205-p24 mAb was produced by transient transfection (calcium phosphate) in 293T cells in serum-free DMEM supplemented with Nutridoma SP (Roche Applied Science). The mAbs were purified on protein G columns (GE Healthcare Bio-Sciences Corp.) and characterized by SDS/PAGE and Western blotting using α-mouse IgG1-HRP (Southern Biotech) or HRP-α-gag-p24 (ImmunoDiagnostics). mAb binding was verified on CHO cells stably transfected with the respective receptor by FACS using phycoerythrin-conjugated goat α-mouse IgG (Jackson ImmunoResearch). Unconjugated α–DEC-205 mAb expressed by stably transfected CHO cells was similarly purified. Recombinant NYVAC-encoding HIV BX08gp120-IIIBGag/Pol/Nef is a replication incompetent vector, kindly provided by Dr. Giuseppe Pantaleo (Centre Hospitalier Universitaire Vaudois (CHUV) Lausanne, Switzerland) and prepared as previously described (75 (link)).
Phycoerythrin conjugated goat α mouse igg
Manufactured by Jackson ImmunoResearch
Phycoerythrin-conjugated goat α-mouse IgG is a secondary antibody reagent that binds to mouse immunoglobulin G (IgG). Phycoerythrin is a fluorescent protein used to label the antibody, allowing for detection and visualization of mouse IgG in various applications.
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2 protocols using phycoerythrin conjugated goat α mouse igg
1
HIV p24 Antigen Expression and Purification
2
HIV p24 Antigen Expression and Purification
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DNA for HIV gag-p24 (aa 133–363 derived from HIV isolate BH10) was cloned in frame into the COOH terminus of the heavy chains of α–mouse-DEC205 as described previously (19 (link)). The fusion α–CD205-p24 mAb was produced by transient transfection (calcium phosphate) in 293T cells in serum-free DMEM supplemented with Nutridoma SP (Roche Applied Science). The mAbs were purified on protein G columns (GE Healthcare Bio-Sciences Corp.) and characterized by SDS/PAGE and Western blotting using α-mouse IgG1-HRP (Southern Biotech) or HRP-α-gag-p24 (ImmunoDiagnostics). mAb binding was verified on CHO cells stably transfected with the respective receptor by FACS using phycoerythrin-conjugated goat α-mouse IgG (Jackson ImmunoResearch). Unconjugated α–DEC-205 mAb expressed by stably transfected CHO cells was similarly purified. Recombinant NYVAC-encoding HIV BX08gp120-IIIBGag/Pol/Nef is a replication incompetent vector, kindly provided by Dr. Giuseppe Pantaleo (Centre Hospitalier Universitaire Vaudois (CHUV) Lausanne, Switzerland) and prepared as previously described (75 (link)).
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