HIV-1Ba-L obtained through the NIH AIDS Research and Reference Reagent Program from Dr. Suzanne Gartner and Dr. Robert Gallo (Gartner et al. 1986 (link)) was grown in primary culture of PBMC. Virus stocks were prepared as previously described (Campbell and Spector 2008 (link)). The 50 % tissue culture infective dose (TCID50) was determined using the Spearman-Karber (Japour et al. 1993 (link)) method using the Alliance HIV-1 p24 antigen ELISA (Perkin Elmer Life Sciences). Cells were infected at a multiplicity of infection (MOI) of 0.1 for 8 h. For virus replication analysis, p24 expression was measured in the culture supernatants by ELISA (Perkin Elmer Life Sciences).
Alliance hiv 1 p24 antigen elisa
Manufactured by PerkinElmer
The Alliance HIV-1 p24 Antigen ELISA is a laboratory equipment product designed for the quantitative detection of HIV-1 p24 antigen. It is an enzyme-linked immunosorbent assay (ELISA) that provides a reliable method for measuring the presence of the HIV-1 p24 protein in biological samples.
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3 protocols using alliance hiv 1 p24 antigen elisa
1
Quantification of HIV-1 Ba-L Replication by p24 ELISA
2
Production and Quantification of HIV-1-GFP Viruses
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HIV-1-GFP single-round infectious viruses were produced by co-transfecting 293T cells with a 1:1:1 ratio of three plasmids: pVSV-G, pCSGW (GFP-reporter) and pCMVΔ8.91 (or pCMVΔ8.91 mutants). Approximately 16 h post-transfection, cells were treated with 10 mM sodium butyrate for 8 h and VLP-containing supernatants were harvested 24 h later. Viral titres were quantified using the Taq-PERT assay to measure viral reverse transcriptase activity [41 (link),42 (link)] or, for reverse transcriptase mutants, the Alliance HIV-1 p24 Antigen ELISA from Perkin-Elmer (NEK050B001KT), following the manufacturer’s instructions. GFP expression was detected on a ZOE fluorescent Cell Imager (BIO-RAD).
3
HIV-1 p24 Antigen ELISA Protocol
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Samples were diluted 1:500 in 0.5% Triton, then p24 protein levels were measured using the Alliance HIV-1 p24 Antigen ELISA (enzyme-linked immunosorbent assay) Kit according to the manufacturer's instructions (PerkinElmer). Plates were read on a Bio-Rad plate reader set to dual wavelength at 405 and 630 nm.
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