resource source identifier antibodies human il 12 p70 antibody  (R&D Systems)

Journal: Experimental Hematology & Oncology

Article Title: Improving CAR-T cell function through a targeted cytokine delivery system utilizing car target-modified extracellular vesicles

doi: 10.1186/s40164-025-00701-z

Figure Lengend Snippet: Manufacture of EVs with functional, surface-displayed IL-12. ( A ) Representative images of IL-12 EVs and control EVs characterized by flow cytometry (independent experiments with n = 3). ( B ) Annexin A1 and IL-12 p70 expression in EVs was detected by western blotting (independent experiments with n = 3). ( C ) The IL-12 concentrations in EVs were quantified via ELISA, and the mean IL-12 concentration of IL-12 EVs was 1887 pg/mL (independent experiments with n = 3). D ) IL-12 EVs were stored at − 80 °C, and their concentration was determined by ELISA analysis at different time points (independent experiments with n = 3). ( E ) Representative dose‒response curves of IFN-γ secretion from T cells (cell density: 1*10 6 /mL) after treatment with doses of rhIL-12 or IL-12 EVs. The EC50 values derived from the graphs were 54.9 pg/mL and 9.4 pg/mL, respectively (independent experiments with n = 3). Data are presented as mean ± SEM. Statistical analysis was performed using unpaired t test. **** p < 0.0001

Article Snippet: For EV molecule characterization, the membranes were incubated overnight at 4 °C with the following antibodies on a shaker: an anti-Annexin A1 antibody (Abcam, ab214486, 1:2000), and anti-IL-12 p70 antibody (R&D Systems, MAB219-SP, 1:1000).

Techniques: Functional Assay, Control, Flow Cytometry, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Concentration Assay, Derivative Assay

Journal: Nature Communications

Article Title: IL-12 drives the expression of the inhibitory receptor NKG2A on human tumor-reactive CD8 T cells

doi: 10.1038/s41467-024-54420-w

Figure Lengend Snippet: Sorted naïve CD8 T cells were activated with CD3/CD28 beads for 24 hours, rested for 2 days and restimulated with CD3/CD28 beads for 8 days in the presence of the indicated cytokines with or without TGF-β. a Data from a representative donor and ( b ) summary of NKG2A expression after 8 days of culture are shown ( n = 5). c Expression of NKG2A by CD8 T cells after stimulation of PBMC with SEB in the presence of isotype control, α-IL-12p70 Ab, α-CD40 Ab or both. Data from one representative donor are shown. d Summary of ( c ) (left) and percentage inhibition of NKG2A expression by CD8 T cells in the different culture conditions (right) ( n = 4). e Detection of IL-12p70 in the culture supernatants of PBMCs stimulated for 3 days with SEB in presence or not of α-CD40 Ab (left) and percentage inhibition of IL-12p70 secretion (right) ( n = 3). b , d , e Data are from distinct healthy donors. Horizontal lines indicate the mean ± SEM. NS= non-significant; p -values were determined by one-way ANOVA with Tukey’s post hoc test ( b , d , g ).

Article Snippet: For IL-12p70 and CD40L blocking experiments, anti-Human Il-12p70 (R&D Systems, cat #MAB219-100, 10 μg/ml final), anti-CD40 (BioLegend, cat #313019, clone HB14, 10 μg/ml final) or isotype control mouse IgG1 (BioLegend, cat #400166, clone MOPC21, 10 μg/ml final) were added in the corresponding wells.

Techniques: Expressing, Control, Inhibition

Journal: Nature Microbiology

Article Title: Post-acute sequelae of SARS-CoV-2 cardiovascular symptoms are associated with trace-level cytokines that affect cardiomyocyte function

doi: 10.1038/s41564-024-01838-z

Figure Lengend Snippet: a , Primary human cardiomyocytes were incubated for 24 h (top row) and 48 h (bottom row) with media alone ((RPMI-1640/B27 with insulin; control) or a cocktail of IL-12, IL-1β, MCP-1 and IL-6 (‘cytokines’). Cytokine concentrations were either ‘Recovered cytokine mimic’ (IL-12, 0.007 fg ml −1 ; IL-1β, 0.01 fg ml −1 ; MCP-1, 0.01 fg ml −1 ; IL-6, 0.004 fg ml −1 ) or ‘PASC-CVS cytokine mimic’ (IL-12, 41 fg ml −1 ; IL-1β, 21 fg ml −1 ; MCP-1, 14 fg ml −1 ; IL-6, 21 fg ml −1 ) to reflect levels detected in the PASC-CVS cohorts. Data are pooled from two independent experiments. Graphs show mean ± s.e.m. Normal distribution of data was assessed with the Shapiro–Wilk test. Statistical significance was determined with a Kruskal–Wallis test with Dunn’s multiple comparison test or Welch ANOVA test and Dunnett’s multiple comparison test. b , Primary human cardiomyocytes were incubated for 24 h (top row) and 48 h (bottom row) with PASC-CVS or Recovered plasma. Data are displayed relative to cardiomyocytes treated with media alone. Mean ± s.e.m. is shown in all graphs. Statistical significance was determined with an ANCOVA adjusted for age, sex and/or site as covariates. Covariates were included in the analysis if statistically significant difference in the covariate was recorded between groups. Each donor is indicated by a unique symbol that is used consistently throughout all figures. Grey horizontal lines indicate the mean value derived from n = 22 Healthy donors. A description of the Healthy donor cohort is presented in Supplementary Table .

Article Snippet: Surface-enhanced Raman scattering nanotags were conjugated with antibodies against IL-1β (MAB601, clone 2805), IL-6 (AF-206, polyclonal), IL-12p70 (MAB611, clone 24945), IL-12p70 (MAB219, clone 24910) or MCP-1 (MAB679, clone 23007) obtained from R&D Systems.

Techniques: Incubation, Control, Comparison, Clinical Proteomics, Derivative Assay

Journal: Nature Microbiology

Article Title: Post-acute sequelae of SARS-CoV-2 cardiovascular symptoms are associated with trace-level cytokines that affect cardiomyocyte function

doi: 10.1038/s41564-024-01838-z

Figure Lengend Snippet: Human cardiomyocytes were incubated for 24 h (top line) or 48 h (bottom line) with media alone ((RPMI-1640/B27 with insulin; control) or individual pro-inflammatory cytokines. Cytokine concentrations were ‘PASC-CVS cytokine mimic’ (IL-12: 41fg/mL; IL-1β 21fg/mL; MCP-1 14fg/mL or IL-6 21fg/mL). Graphs show mean ± SEM. Normal distribution of data was assessed with the Shapiro-Wilk test. Mean ± SEM is shown. Statistical significance was determined with a Kruskal-Wallis test with Dunn’s multiple comparison test or Welch ANOVA test and Dunnett’s multiple comparison test. Data is pooled from three independent experiments.

Article Snippet: Surface-enhanced Raman scattering nanotags were conjugated with antibodies against IL-1β (MAB601, clone 2805), IL-6 (AF-206, polyclonal), IL-12p70 (MAB611, clone 24945), IL-12p70 (MAB219, clone 24910) or MCP-1 (MAB679, clone 23007) obtained from R&D Systems.

Techniques: Incubation, Control, Comparison