Retro tek siv p27 antigen elisa kit

Jurkat cells were seeded at 1 × 10⁶ cells/mL for 24 h, and infected with known amounts (10 ng/mL p24/p27 units per 10⁶ cells) of HIV-1 (MN) [36 (link),37 (link)] and HIV-2 (ROD) and cultured for different days as indicated. PBMC cells were infected with known amounts (5 ng/mL p24/p27 units per 10⁶ cells) of HIV-1 (MN) and HIV-2 (ROD). Two hours post-infection, tubes were washed with PBS to remove un-adsorbed virus and cells were fed with complete RPMI (RPMI 1640, 10% FBS, 1% penicillin–streptomycin for Jurkat cells and supplemented with IL-2 for PBMC) and cultured in T75 flask. The cells were harvested at different time points as indicated. HIV-1 and HIV-2 replication was quantitated by Alliance HIV-1 p24 Antigen ELISA Kit (Perkin Elmer, Waltham, MA, USA) and RETRO-TEK SIV p27 Antigen ELISA Kit (ZeptoMetrix, Buffalo, NY, USA).

PBMC cells were infected with known amounts (5 ng/mL HIV-1 p24 units per 10⁶ cells) of T-tropic HIV-1 (MN) and adherent MDMs were infected with equivalent amounts of (5 ng/mL HIV-1 p24 units/10⁶ cells of M-tropic HIV-1 Ba-L, Cat# 510, NIH AIDS Reagent, Bethesda, MD, USA) viruses. Both the cells were infected with equivalent amounts of dual tropic HIV-2 (ROD, 5 ng/mL SIV p27 units/million cells, Cat# 0121, NIBSC AIDS Reagent Program). After two hours post-infection, unadsorbed viruses were removed by washing the cells with PBS and cultured in complete media until further use. The cells were harvested on day 7 post infection as indicated. HIV-1 and HIV-2 replication was quantitated by an Alliance HIV-1 p24 Antigen ELISA Kit (Perkin Elmer, Waltham, MA, USA) and a RETRO-TEK SIV p27 Antigen ELISA Kit (ZeptoMetrix, Buffalo, NY, USA).