VOAs were performed using total and/or resting CD4+ T-cells as previously described [19 (link)]. Each VOA used 6 million to 13 million CD4+ T-cells seeded in 3-fold serial dilutions with six replicates per dilution, beginning with 1 million cells per well. Wells that were positive for p24 by ELISA (Alliance HIV-1 P24 Antigen ELISA Kit, Perkin Elmer) on days 14 or 21 had their supernatants harvested and stored at -80°C for subsequent single-genome sequencing. Maximum likelihood estimates were applied to calculate infectious units per million cells (IUPM) [44 (link)].
Alliance hiv 1 p24 antigen elisa kit
Manufactured by PerkinElmer
Sourced in United States
The Alliance HIV-1 p24 Antigen ELISA kit is a laboratory equipment product designed to detect the presence of the HIV-1 p24 antigen in biological samples. It utilizes the enzyme-linked immunosorbent assay (ELISA) technique to perform the analysis.
Automatically generated - may contain errors
Lab products found in correlation
Retro tek siv p27 antigen elisa kit, by ZeptoMetrix (2 mentions)
Cd4 t cell isolation kit 2, by Miltenyi Biotec (1 mentions)
Dynabead human t activator cd3 cd28, by Thermo Fisher Scientific (1 mentions)
Quikchange site directed mutagenesis kit, by Agilent Technologies (1 mentions)
Facscalibur, by BD (1 mentions)
Trypsin edta, by Merck Group (1 mentions)
Q5 polymerase, by Thermo Fisher Scientific (1 mentions)
Pspaxii, by Addgene (1 mentions)
Versamax, by Molecular Devices (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
High glucose dmem, by Thermo Fisher Scientific (1 mentions)
Galacton star substrate, by Thermo Fisher Scientific (1 mentions)
Hek293t, by American Type Culture Collection (1 mentions)
Rnase free dnase 1, by Roche (1 mentions)
Sw28 rotor, by Beckman Coulter (1 mentions)
Polybrene, by Merck Group (1 mentions)
Ifn γ, by R&D Systems (1 mentions)
Interleukin 4 (il 4), by R&D Systems (1 mentions)
26 protocols using alliance hiv 1 p24 antigen elisa kit
1
Quantifying HIV-1 Latent Reservoir
2
GHOST Cell HIV-1 Infection Kinetics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
GHOST cells (NIH AIDS Reagent Program, Division of AIDS, National Institute of Allergy and Infectious Diseases, National Institutes of Health) were seeded at 2×105 per well in 12-well plates the day before infection. Equal amount of each virus (5 ng p24) was added to each well. After absorption for 6 h at 37°C, the cells were washed three times and resupplemented with complete medium. The cell culture was maintained for 10 days. Cell culture supernatants were collected every two days to monitor viral replication kinetics by determining the p24 concentrations using the Alliance HIV-1 p24 Antigen ELISA kit (PerkinElmer, Waltham, MA, USA).
3
HIV-1 and HIV-2 Infection Kinetics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
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).
4
Anti-HIV-1 Activity of 72B Extract
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PM-1 cells were used to evaluate the anti-HIV-1 BaL activity of 72B extract.Quantitation of the p24 Gag protein present in supernatants of HIV-1 BaL infected PM-1 cells was assessed using the Alliance HIV-1 P24 ANTIGEN ELISA Kit (Perkin Elmer, Waltham, Massachusetts, USA), according to the manufacturer’s protocol, on culture supernatant using the antigen-capture enzyme-linked immunosorbent assay test (ELISA) (Perkin Elmer).
5
Isolation and Activation of CD4+ T Cells
6
HIV-1 Inhibition by RNA Treatments
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CCRF-CEM cells were infected with HIV-1 IIIB for 5 days (MOI 0.001). Prior to RNA treatments the infected cells were gently washed with PBS three times to remove free virus. On the day of the experiments (day 1), 2.5×105 infected cells and 2.5×105 uninfected cells were mixed and then incubated with refolded experimental RNAs at 800 nM final concentration in 12-well plates at 37°C at day 1 and day 3 (twice treatment), which were accompanied by addition of the buffer (mock) or Trichostatin A (TSA) at 1 µM final concentration or 5-Aza-2'-deoxycytidine (5' AzaC) at 4 µM final concentration. The culture supernatants were collected at day 7. As control, irrelative aptamer-siRNA chimera (BAFF-R aptamer R-1-LTR-362 DsiRNA chimera) and A-1 aptamer-scrambled siRNA chimera were used here. The HIV-1 p24 antigen analyses were performed using an Alliance HIV-1 p24 Antigen ELISA kit (PerkinElmer, CA) according to the manufacturer's instructions.
7
HIV-1 Inhibition in Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CCRF-CEM cells or human PBMCs-CD4+ cells were infected with HIV-1 IIIB or NL4-3 for 5 days (MOI 0.001). HIV-1 LAV-infected Jurkat E6 Cells (J1.1) were gently washed with 1 mL PBS three times to remove free viruses and then cultured one day for HIV-1 induction. Prior to RNA treatments the infected cells were gently washed with PBS three times to remove free virus. Next 1×106 infected cells and 1×106 uninfected cells were incubated with formulated RNA conjugates at 800 nM final concentration in 24-well plates at 37°C. The formulated RNA conjugates were added at days 1, 3 and 5. The culture supernatants were collected at different times (7 d and 10 d). The p24 antigen analyses were performed using a HIV-1 p24 Antigen Assay (Alliance HIV-1 p24 Antigen ELISA kit, PerkinElmer, CA) according to the manufacturer's instructions.
8
Generating HIV-1 Viral Constructs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HIV-1YU2 and HIV-1YU2TM2 were produced by transiently transfecting HEK 293T/17 cells using a construct consisting of HIV-1NL4/3 backbone carrying the HIV-1YU2 envelope (Zhang et al., 2002 (link)). HIV-1YU2TM2 harbors the mutations N160K, N332K, and G458D that were introduced by site-directed mutagenesis using the QuikChange Site-Directed Mutagenesis kit (Agilent Technologies). The concentration of the virus was determined by measuring p24 using the Alliance HIV-1 p24 Antigen ELISA kit (PerkinElmer). Humanized mice were infected by intraperitoneal injection of HIV-1YU2 or HIV-1YU2TM2 viral supernatant containing 110 ng of p24.
9
HIV-1 Infection Assay in Jurkat Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Direct infection assays were performed utilizing Jurkat-R5-control and Jurkat-R5-Gal3 cells (1 × 105/well). Briefly, HIV-1 NL4-3 or CRF07_BC viruses (MOI = 0.1–0.01) were subjected to incubation with cells for 2 h at 37 °C in serum-free medium, followed by rinsing in phosphate-Buffered Saline (PBS). Subsequently, 2 mL of Roswell Park Memorial Institute Medium (RPMI) or Dulbecco’s Modified Eagle Medium (DMEM) medium containing 10% serum, antibiotics, and polybrene (8 μg/mL) were added, followed by incubation in 5% CO2 at 37 °C. The 100 μL supernatants were collected and refilled with 100 μL fresh culture medium in each well every two days. Collected viral supernatants were subjected to HIV-1 p24 quantification using Alliance HIV-1 P24 Antigen ELISA Kit (PerkinElmer; Cat. No. NEK050B, Waltham, MA, USA).
10
HIV-1 Infection Kinetics Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Direct infection assays were preformed utilizing Jurkat-R5-control and Jurkat-R5-Gal3 cells (1 × 105 /well). Briefly, HIV-1 NL4-3 or CRF07_BC viruses (MOI = 0.1–0.01) were subjected to incubation with cells for 2 h at 37 °C in serum-free medium, followed by rinsing in phosphate-buffered saline (PBS). Subsequently, 2 mL of RPMI or DMEM medium containing 10% serum, antibiotics, and polybrene (8 μg/mL) were added, followed by incubation in 5% CO2 at 37 °C. A total of 100 μL supernatants were collected and refilled with 100 μl fresh culture medium in each well in every two days. Collected viral supernatants were subjected to HIV-1 p24 quantification using the Alliance HIV-1 P24 Antigen ELISA Kit (PerkinElmer Life and Analytical Sciences, Boston, MA; Cat. No. NEK050B).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!