Description
• Measures the p24 core protein of recombinant HIV-1 based lentivirus
• Lentivirus quantitation is performed on a standard microplate reader
• p24 Standard included
Quantitation of the HIV-1 p24 core protein of a lentiviral vector is an efficient, well-published method for determining lentivirus physical titer. The QuickTiter Lentivirus Quantitation Kits provide a convenient method of p24 antigen quantitation.
Introduction: Lentivirus vector based on the human immunodeficiency virus-1 (HIV-1) has become a promising vector for gene transfer studies. The advantageous feature of lentivirus vector is the ability of gene transfer and integration into dividing and non-dividing cells1-2. The pseudotyped envelope with vesicular stomatitis virus envelope G (VSV-G) protein broadens the target cell range. Lentiviral vectors have been shown to deliver genes to neurons, lymphocytes and macrophages, cell types that previous retrovirus vectors could not be used. Lentiviral vectors have also proven to be effective in transducing brain, liver, muscle, and retina in vivo without toxicity or immune responses. Recently, the lentivirus system is widely used to integrate siRNA efficiently in a wide variety of cell lines and primary cells both in vitro and in vivo. Lentivirus particles are produced from 293T cells through transient transfection of 3 or 4 plasmids that encodes for the components of the virion. Viral medium containing viral particles produced by packaging cells within 48-72 hr can be harvested. To ensure that pseudoviral medium is viable, and to control the number of copies of integrated viral constructs per target cell, the viral titer needs to be determined before proceeding with transduction experiments. Viral titer can be determined by transduction of HT-1080 or HeLa cells, and followed by antibiotic selection of stable clones. However, it takes weeks to generate sizable stable cell colonies for counting and calculating the titer results. MyBioSource's QuickTiter HIV Lentiviral Quantitation Kit (HIV p24 ELISA) is an enzyme immunoassay developed for detection and quantitation of the HIV-1 p24 core protein. A mouse monoclonal antibody to HIV-1 p24 is coated onto strip wells of microtiter plate. The quantity of HIV p24 antigen is determined by comparing its absorbance with that of known recombinant p24 antigen standard curve. The kit has a detection sensitivity limit of 1 ng/mL HIV p24, or about 10,000 to 100,000 TU/mL VSVG-pseudotyped lentivirus samples3-5. Each kit provides sufficient reagents to perform up to 96 assays including standard curve and unknown samples. The kit is suitable for both viral supernatant and purified virus. The QuickTiter HIV Lentiviral Quantitation Kit is intended for research use only, and not for diagnostic applications