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    Sino Biological hiv p24 elisa
    STAT1 or STAT3 inhibitors suppress <t>HIV-1</t> infection of MDM. MDM were pretreated with 100 nM Fludarabine (STAT1 inhibitor) or 50 nM LLL12 (STAT3 inhibitor) 24 h before infection and on PID 1, 4, and 7. ELISA was used to measure supernatant <t>p24</t> on PID 4, 7, and 10 in cultures treated with (A) Fludarabine and (B) LLL12. Bars represent mean ± SEM; * p
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    STAT1 or STAT3 inhibitors suppress HIV-1 infection of MDM. MDM were pretreated with 100 nM Fludarabine (STAT1 inhibitor) or 50 nM LLL12 (STAT3 inhibitor) 24 h before infection and on PID 1, 4, and 7. ELISA was used to measure supernatant p24 on PID 4, 7, and 10 in cultures treated with (A) Fludarabine and (B) LLL12. Bars represent mean ± SEM; * p

    Journal: AIDS Research and Human Retroviruses

    Article Title: HIV-1 Infection Primes Macrophages Through STAT Signaling to Promote Enhanced Inflammation and Viral Replication

    doi: 10.1089/aid.2016.0273

    Figure Lengend Snippet: STAT1 or STAT3 inhibitors suppress HIV-1 infection of MDM. MDM were pretreated with 100 nM Fludarabine (STAT1 inhibitor) or 50 nM LLL12 (STAT3 inhibitor) 24 h before infection and on PID 1, 4, and 7. ELISA was used to measure supernatant p24 on PID 4, 7, and 10 in cultures treated with (A) Fludarabine and (B) LLL12. Bars represent mean ± SEM; * p

    Article Snippet: HIV-1 Gag p24 in culture supernatants was measured by ELISA (Sino Biological, Daxing, China).

    Techniques: Infection, Enzyme-linked Immunosorbent Assay

    STAT1 and STAT3 are involved in HIV-1-induced priming. MDM were treated with 100 nM Fludarabine (STAT1 inhibitor) or 50 nM LLL12 (STAT3 inhibitor) 24 h before infection and on PID 1, 4, and 7. Ten days postinfection, cells were activated with 1 μg/mL CL097 for 6 h. Intracellular p24 and TNF were measured by flow cytometry. Bars represent mean ± SEM MFI of TNF in uninfected cells and infected cells gated on either p24-negative or p24-positive populations; * p

    Journal: AIDS Research and Human Retroviruses

    Article Title: HIV-1 Infection Primes Macrophages Through STAT Signaling to Promote Enhanced Inflammation and Viral Replication

    doi: 10.1089/aid.2016.0273

    Figure Lengend Snippet: STAT1 and STAT3 are involved in HIV-1-induced priming. MDM were treated with 100 nM Fludarabine (STAT1 inhibitor) or 50 nM LLL12 (STAT3 inhibitor) 24 h before infection and on PID 1, 4, and 7. Ten days postinfection, cells were activated with 1 μg/mL CL097 for 6 h. Intracellular p24 and TNF were measured by flow cytometry. Bars represent mean ± SEM MFI of TNF in uninfected cells and infected cells gated on either p24-negative or p24-positive populations; * p

    Article Snippet: HIV-1 Gag p24 in culture supernatants was measured by ELISA (Sino Biological, Daxing, China).

    Techniques: Infection, Flow Cytometry

    IFNγ- and HIV-1-induced macrophage-priming phenotype. (A) A panel of 17 macrophage activation genes was measured by RT-qPCR and fold change over unactivated MDM (medium only) was used to perform hierarchical clustering between the treatment groups. (B) Gene expression was compared between HIV-1-induced and IFNγ-induced MDM priming. The solid line represents the linear regression of all genes ( r = .679, P = .003). Gene expression was compared between MDM (C) activated with LPS alone (2 h) and MDM primed with IFNγ and then activated with LPS (2 h) or (D) activated with LPS alone (2 h) and MDM infected with 500 TCID 50 HIV-1 AD8 and then activated with LPS (2 h). The red circles indicate genes modified in a similar direction (up or down) by both IFNγ and HIV-1 compared with LPS alone. The blue circles indicate genes enhanced by IFNγ, but suppressed or unaffected (ND, no difference) by HIV-1 infection compared with LPS alone. The fold change for each gene in comparison with LPS alone is indicated in parentheses . Experiment was conducted in three donors. RT-qPCR, reverse transcriptase quantitative polymerase chain reaction; TCID 50

    Journal: AIDS Research and Human Retroviruses

    Article Title: HIV-1 Infection Primes Macrophages Through STAT Signaling to Promote Enhanced Inflammation and Viral Replication

    doi: 10.1089/aid.2016.0273

    Figure Lengend Snippet: IFNγ- and HIV-1-induced macrophage-priming phenotype. (A) A panel of 17 macrophage activation genes was measured by RT-qPCR and fold change over unactivated MDM (medium only) was used to perform hierarchical clustering between the treatment groups. (B) Gene expression was compared between HIV-1-induced and IFNγ-induced MDM priming. The solid line represents the linear regression of all genes ( r = .679, P = .003). Gene expression was compared between MDM (C) activated with LPS alone (2 h) and MDM primed with IFNγ and then activated with LPS (2 h) or (D) activated with LPS alone (2 h) and MDM infected with 500 TCID 50 HIV-1 AD8 and then activated with LPS (2 h). The red circles indicate genes modified in a similar direction (up or down) by both IFNγ and HIV-1 compared with LPS alone. The blue circles indicate genes enhanced by IFNγ, but suppressed or unaffected (ND, no difference) by HIV-1 infection compared with LPS alone. The fold change for each gene in comparison with LPS alone is indicated in parentheses . Experiment was conducted in three donors. RT-qPCR, reverse transcriptase quantitative polymerase chain reaction; TCID 50

    Article Snippet: HIV-1 Gag p24 in culture supernatants was measured by ELISA (Sino Biological, Daxing, China).

    Techniques: Activation Assay, Quantitative RT-PCR, Expressing, Infection, Modification, Real-time Polymerase Chain Reaction

    HIV-1 infection directly primes MDM for hyperresponsiveness to TLR4 and TLR7/8-mediated activation. The effects of IFNγ stimulation versus HIV-1 infection on LPS- and CL097-induced TNF production were compared in MDM. (A) Untreated (medium alone), IFNγ-primed (10 ng/mL for 18 h), or HIV-1 infected MDM (50–5,000 TCID 50 for 7 days) were left untreated or activated with 10 ng/mL LPS for 6 h. ELISA was used to measure supernatant TNF. Bars represent the mean ± SEM * p

    Journal: AIDS Research and Human Retroviruses

    Article Title: HIV-1 Infection Primes Macrophages Through STAT Signaling to Promote Enhanced Inflammation and Viral Replication

    doi: 10.1089/aid.2016.0273

    Figure Lengend Snippet: HIV-1 infection directly primes MDM for hyperresponsiveness to TLR4 and TLR7/8-mediated activation. The effects of IFNγ stimulation versus HIV-1 infection on LPS- and CL097-induced TNF production were compared in MDM. (A) Untreated (medium alone), IFNγ-primed (10 ng/mL for 18 h), or HIV-1 infected MDM (50–5,000 TCID 50 for 7 days) were left untreated or activated with 10 ng/mL LPS for 6 h. ELISA was used to measure supernatant TNF. Bars represent the mean ± SEM * p

    Article Snippet: HIV-1 Gag p24 in culture supernatants was measured by ELISA (Sino Biological, Daxing, China).

    Techniques: Infection, Activation Assay, Enzyme-linked Immunosorbent Assay

    Only productively infected (p24 + ) MDM are primed. (A) MDM were infected with 5000 TCID 50 HIV-1 NL4-3-BaL-HSA encoding the murine heat-stable antigen (HSA; CD24), which is expressed on the surface of productively infected cells. CD24 + and CD24 − cells were sorted using magnetic beads and expression of HIV-1 gag p24 was measured by western blot to confirm efficient sorting of infected cells. (B) Uninfected and infected (CD24 − and CD24 + ) MDM were replated and stimulated with 10 ng/mL LPS for 6 h. ELISA measured secreted TNF. The experiment was conducted in one donor. ND, not detectable.

    Journal: AIDS Research and Human Retroviruses

    Article Title: HIV-1 Infection Primes Macrophages Through STAT Signaling to Promote Enhanced Inflammation and Viral Replication

    doi: 10.1089/aid.2016.0273

    Figure Lengend Snippet: Only productively infected (p24 + ) MDM are primed. (A) MDM were infected with 5000 TCID 50 HIV-1 NL4-3-BaL-HSA encoding the murine heat-stable antigen (HSA; CD24), which is expressed on the surface of productively infected cells. CD24 + and CD24 − cells were sorted using magnetic beads and expression of HIV-1 gag p24 was measured by western blot to confirm efficient sorting of infected cells. (B) Uninfected and infected (CD24 − and CD24 + ) MDM were replated and stimulated with 10 ng/mL LPS for 6 h. ELISA measured secreted TNF. The experiment was conducted in one donor. ND, not detectable.

    Article Snippet: HIV-1 Gag p24 in culture supernatants was measured by ELISA (Sino Biological, Daxing, China).

    Techniques: Infection, Magnetic Beads, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

    STAT1 and STAT3 are involved in HIV-1 replication. MDM were treated with 100 nM Fludarabine (STAT1 inhibitor) or 50 nM LLL12 (STAT3 inhibitor) 24 h before infection and total DNA was extracted on PID 1. Total (A) or integrated (B) HIV-1 proviral genomes were measured using qPCR, normalized to ApoB , and compared with medium alone (Fludarabine control) or DMSO (LLL12 control). Bars represent the mean ± SEM. (C) MDM treated with Fludarabine or LLL12, 24 h pre- and postinfection, with single-cycle luciferase reporter HIV-1 JRFL were lysed on PID 4 and luciferase activity was measured. Bars represent mean ± SEM relative light units (RLU); ** p

    Journal: AIDS Research and Human Retroviruses

    Article Title: HIV-1 Infection Primes Macrophages Through STAT Signaling to Promote Enhanced Inflammation and Viral Replication

    doi: 10.1089/aid.2016.0273

    Figure Lengend Snippet: STAT1 and STAT3 are involved in HIV-1 replication. MDM were treated with 100 nM Fludarabine (STAT1 inhibitor) or 50 nM LLL12 (STAT3 inhibitor) 24 h before infection and total DNA was extracted on PID 1. Total (A) or integrated (B) HIV-1 proviral genomes were measured using qPCR, normalized to ApoB , and compared with medium alone (Fludarabine control) or DMSO (LLL12 control). Bars represent the mean ± SEM. (C) MDM treated with Fludarabine or LLL12, 24 h pre- and postinfection, with single-cycle luciferase reporter HIV-1 JRFL were lysed on PID 4 and luciferase activity was measured. Bars represent mean ± SEM relative light units (RLU); ** p

    Article Snippet: HIV-1 Gag p24 in culture supernatants was measured by ELISA (Sino Biological, Daxing, China).

    Techniques: Infection, Real-time Polymerase Chain Reaction, Luciferase, Activity Assay

    Type I IFN restricts HIV-1 replication. (A) TZM-bl cells were plated in 96-well plates (10,000 cells/well) and allowed to adhere overnight. The following day, the cells were pre-treated with IFN-β for 24 hours and then infected with 100 TCID 50 HIV HSA . After 48 hours, the amount of HIV-1 replication was measured by luciferase assay. The dashed line indicates the level of luciferase activity in cells that were not pretreated with IFN. (B) Monocytes (100,000 cells/well) were differentiated into MDMs in M-CSF and then pretreated with IFN-β for 24 hours followed by infection with 150 TCID 50 HIV HSA for 7 days. Supernatant Gag p24 was measured by ELISA (n = 3 donors). Data were normalized to Gag p24 levels of infected cells with no IFN-β treatment (100% replication) and no infection (0% infection) for each donor. A nonlinear regression was plotting of the combined data with GraphPad Prism (v 7.02) use the least squares fit method to determine the IC 50 .

    Journal: Journal of leukocyte biology

    Article Title: CRISPR/Cas9 knockout of USP18 enhances type I IFN responsiveness and restricts HIV-1 infection in macrophages

    doi: 10.1002/JLB.3MIA0917-352R

    Figure Lengend Snippet: Type I IFN restricts HIV-1 replication. (A) TZM-bl cells were plated in 96-well plates (10,000 cells/well) and allowed to adhere overnight. The following day, the cells were pre-treated with IFN-β for 24 hours and then infected with 100 TCID 50 HIV HSA . After 48 hours, the amount of HIV-1 replication was measured by luciferase assay. The dashed line indicates the level of luciferase activity in cells that were not pretreated with IFN. (B) Monocytes (100,000 cells/well) were differentiated into MDMs in M-CSF and then pretreated with IFN-β for 24 hours followed by infection with 150 TCID 50 HIV HSA for 7 days. Supernatant Gag p24 was measured by ELISA (n = 3 donors). Data were normalized to Gag p24 levels of infected cells with no IFN-β treatment (100% replication) and no infection (0% infection) for each donor. A nonlinear regression was plotting of the combined data with GraphPad Prism (v 7.02) use the least squares fit method to determine the IC 50 .

    Article Snippet: HIV-1 Gagp24 was measured in cell-free supernatants lysed with 0.5% Triton X-100 in PBS by HIV-1 Gagp24 ELISA kit according to the manufacturer’s instructions (Sino Biological, Beijing, China).

    Techniques: Infection, Luciferase, Activity Assay, Enzyme-linked Immunosorbent Assay

    shRNA knockdown of USP18 inhibits HIV-1 replication in THP-1 cells. THP-1 cells expressing control or USP18–5 shRNA were activated with PMA. (A) THP-1 cells were infected with 5000 TCID 50 HIV AD for 5 days and supernatant Gag p24 was measured by ELISA. (B) THP-1 cells were infected with 5000 TCID 50 HIV HSA for 5 days and intracellular Gag p24 expression was measured by Western blot. A ratio paired T-test was used to compare supernatant Gag p24 levels between control shRNA and USP18–5 shRNA expressing cells. N.D. = not detected, * p

    Journal: Journal of leukocyte biology

    Article Title: CRISPR/Cas9 knockout of USP18 enhances type I IFN responsiveness and restricts HIV-1 infection in macrophages

    doi: 10.1002/JLB.3MIA0917-352R

    Figure Lengend Snippet: shRNA knockdown of USP18 inhibits HIV-1 replication in THP-1 cells. THP-1 cells expressing control or USP18–5 shRNA were activated with PMA. (A) THP-1 cells were infected with 5000 TCID 50 HIV AD for 5 days and supernatant Gag p24 was measured by ELISA. (B) THP-1 cells were infected with 5000 TCID 50 HIV HSA for 5 days and intracellular Gag p24 expression was measured by Western blot. A ratio paired T-test was used to compare supernatant Gag p24 levels between control shRNA and USP18–5 shRNA expressing cells. N.D. = not detected, * p

    Article Snippet: HIV-1 Gagp24 was measured in cell-free supernatants lysed with 0.5% Triton X-100 in PBS by HIV-1 Gagp24 ELISA kit according to the manufacturer’s instructions (Sino Biological, Beijing, China).

    Techniques: shRNA, Expressing, Infection, Enzyme-linked Immunosorbent Assay, Western Blot

    IFNAR blocking inhibits HIV-1 replication and blocks HIV-1-induced USP18 expression in MDMs. MDMs were infected with 1000 TCID 50 HIV AD in the presence of α-IFNAR2 neutralizing antibody or IgG2a isotype control antibody. Supernatants were harvested on days 4 and 8 post-infection and supernatant Gag p24 was measured by ELISA. Protein lysates were collected on day 4 post-infection and Gag p24 and USP18 expression were assessed by Western blot.

    Journal: Journal of leukocyte biology

    Article Title: CRISPR/Cas9 knockout of USP18 enhances type I IFN responsiveness and restricts HIV-1 infection in macrophages

    doi: 10.1002/JLB.3MIA0917-352R

    Figure Lengend Snippet: IFNAR blocking inhibits HIV-1 replication and blocks HIV-1-induced USP18 expression in MDMs. MDMs were infected with 1000 TCID 50 HIV AD in the presence of α-IFNAR2 neutralizing antibody or IgG2a isotype control antibody. Supernatants were harvested on days 4 and 8 post-infection and supernatant Gag p24 was measured by ELISA. Protein lysates were collected on day 4 post-infection and Gag p24 and USP18 expression were assessed by Western blot.

    Article Snippet: HIV-1 Gagp24 was measured in cell-free supernatants lysed with 0.5% Triton X-100 in PBS by HIV-1 Gagp24 ELISA kit according to the manufacturer’s instructions (Sino Biological, Beijing, China).

    Techniques: Blocking Assay, Expressing, Infection, Enzyme-linked Immunosorbent Assay, Western Blot

    iMacs support HIV-1 replication. iMacs were infected with 1000 TCID 50 HIV HSA for 8 days. (A) Supernatant Gag p24 was measured by ELISA. (B) Intracellular Gag p24 and USP18 expression was measured by Western blot. (C) USP18 +/+ and USP18 −/− iMacs were infected with 1000 TCID 50 HIV HSA for 8 days. Supernatant Gag p24 was measured by ELISA (n=5). (D) Intracellular Gag p24 and USP18 were measured by Western blot. Representative blot of three experiments is shown. A paired ratio T-test was used to determine statistical significance. ** p

    Journal: Journal of leukocyte biology

    Article Title: CRISPR/Cas9 knockout of USP18 enhances type I IFN responsiveness and restricts HIV-1 infection in macrophages

    doi: 10.1002/JLB.3MIA0917-352R

    Figure Lengend Snippet: iMacs support HIV-1 replication. iMacs were infected with 1000 TCID 50 HIV HSA for 8 days. (A) Supernatant Gag p24 was measured by ELISA. (B) Intracellular Gag p24 and USP18 expression was measured by Western blot. (C) USP18 +/+ and USP18 −/− iMacs were infected with 1000 TCID 50 HIV HSA for 8 days. Supernatant Gag p24 was measured by ELISA (n=5). (D) Intracellular Gag p24 and USP18 were measured by Western blot. Representative blot of three experiments is shown. A paired ratio T-test was used to determine statistical significance. ** p

    Article Snippet: HIV-1 Gagp24 was measured in cell-free supernatants lysed with 0.5% Triton X-100 in PBS by HIV-1 Gagp24 ELISA kit according to the manufacturer’s instructions (Sino Biological, Beijing, China).

    Techniques: Infection, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot