QuickTiter™ HIV Lentivirus Quantitation Kit (p24 ELISA)

Product Description

• Measures the p24 core protein of recombinant HIV-1-based lentivirus
• Lentivirus quantitation is performed on a standard microplate reader
• p24 Standard included

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.

The 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.

HIV p24 ELISA Standard Curve

p24 Titer of GFP Lentiviral Supernatant

A GFP lentiviral construct was cotransfected with a packaging mix into 293AD cells (Cat.# AD-100). The conditioned medium was harvested 48 hrs after transfection. GFP expression was shown in HEK293 cells infected with the GFP lentiviral supernatant for 3 days. The p24 level in the diluted lentiviral supernatant (1:10 dilution) was determined as described in the assay instructions.