The Lenti-X p24 Rapid Titer Kit uses a standard ELISA to quickly determine the titer of any HIV-1-based lentiviral supernatant. The wells of the included microtiter plate (12 x 8-well strips) are coated with anti-p24 capture antibody, which quantitatively binds the p24 virus core/capsid protein in your test samples. Specifically bound p24 is detected in a typical sandwich ELISA format using a biotinylated anti-p24 secondary antibody, a streptavidin-HRP conjugate, and a color-producing substrate. A p24 standard curve quantifies p24 content, which allows the lentivirus titer to be calculated.
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Rapid lentivirus titration by p24 ELISA
What is p24?
Recombinant lentiviruses generated for research use, whether they are 3rd- or 4th-generation (e.g., generated with Lenti-X kits), contain major structural proteins, including the VSV-G or ecotropic envelope proteins, matrix proteins, and virus core proteins (Figure 2). The gag gene encodes the viral capsid protein (p24), the nucleocapsid proteins (p6 and p7), and a matrix protein (p17). The Lenti-X p24 Rapid Titer Kit specifically measures the amount of p24 capsid protein present in your viral supernatant. The level of p24 correlates directly with virus titer.
How does it work?
The virus in your lentiviral supernatant is first lysed in the supplied buffer system to release p24. The sample is then applied to a 96-well microtitration plate (made up of 12 separable 8-well strips), in which the wells are precoated with murine anti-p24 capture antibody. After washing to remove unbound lysate, the amount of specifically bound p24 is then detected using a biotinylated anti-p24 secondary antibody, streptavidin-HRP (horseradish peroxidase), and a color development reagent. The kit includes a p24 control which is used to generate a standard curve and calibrate the p24 equivalent of your supernatant and its titer.
The titer of your supernatant is determined by correlating its p24 value to virus content or infectivity (Figure 3). Once the relationship is defined, your virus production system can be calibrated to determine a relationship between p24 levels and infectivity. Alternatively, you can calculate the amount of virus based on p24 content as described in the user manual.
As a leader in lentiviral technology, we offer a full range of Lenti-X expression systems and accessories for the effective delivery of cDNAs or shRNAs into any cell type using high-titer lentiviral vectors.
Lenti-X Concentrator video
Lenti-X Concentrator is a simple and fast method for concentrating any lentiviral stock—just mix your lentiviral supernatant with the concentration reagent, incubate, and spin the mixture in a standard centrifuge. Watch the video to see how it works.Watch Concentrator video Lentiviral packaging overview
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