HIV-1 Gag p24 Quantikine ELISA Kit Summary

Product Summary

The Quantikine^® HIV-1 Gag p24 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure HIV-1 Gag p24 in cell culture supernates.

Preparation and Storage

Background: HIV-1 Gag p24

The gag gene of human immunodeficiency virus 1 (HIV-1) encodes a precursor protein known as Pr55^Gag. The viral protease PR cleaves this precursor to generate p17, p24, p7, and p6 proteins which are required for virus particle assembly. p24 is a major viral core structural protein. Its measurement is commonly used as an indicator of HIV-1 infection and viral load.

Assay Procedure

Refer to the product datasheet for the complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.

1.     Prepare all reagents, standard dilutions, and samples as directed in the product insert.

2.     Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

3.     Add 50 μL of Assay Diluent to each well.

4.     Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

5.     Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6.     Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7.     Aspirate and wash 5 times.

8.     Add 100 μL Substrate Solution to each well.

9.     Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.