human il 1β il 1f2 duoset elisa kit  (R&D Systems)

Journal: Nature chemical biology

Article Title: A druggable redox switch on SHP1 controls macrophage inflammation

doi: 10.1038/s41589-026-02163-8

Figure Lengend Snippet: a, LPS-induced (100 ng/ml) SHP1 Tyr536, Tyr564 and Ser591 phosphorylation in iBMDMs over a time course of 2 h in iBMDMs pre-treated with DMSO or SCA1 (50 μM) for 3 h. Immunoblots shown are representative of three independent experiments. b , LPS-induced (100 ng/ml) STAT3 Tyr705 and Ser727 phosphorylation in iBMDMs over a time course of 2 h in iBMDMs pre-treated with DMSO or SCA1 (50 μM) for 3 h. Immunoblots shown are representative of three independent experiments. c,e,f , Pro-inflammatory cytokine IL-6 and TNF levels in cell supernatants of iBMDMs pre-incubated 3 h with SCA1, SCA1-NC or the respective derivatives over a dose response (0.313–80 μM) followed by 6 h LPS stimulation (100 ng/ml) ( n = 4). For measurement of pro-inflammatory cytokine IL-1β levels in cell supernatants, iBMDMs were pre-treated with LPS (100 ng/ml) for 3 h followed by treatment with DMSO or SCA9 at the indicated concentrations for 45 min and primed with adenosine triphosphate (ATP, 5 mM) for 45 min ( n = 3). Absolute cytokine production as a function of concentration is used to calculate IC 50 values. d , Pro-inflammatory cytokine IL-6 and TNF levels in cell supernatants of iBMDMs treated 3 h with SCA1 (20 μM) alone or 6 h with LPS (100 ng/ml) alone ( n = 3). g, Phagocytic activity of iBMDMs pre-treated 3 h with SCA1, SCA9, SCA7 or SCA25 (10 μM) followed by 6 h LPS stimulation (100 ng/ml) ( n = 6). h, Relative fold changes in protein abundance between iBMDMs treated with SCA1 (50 μM) for 3 h followed by LPS (100 ng/ml, 15 min) versus LPS alone ( n = 3). Data are mean ± s.e.m. (in d,h ) or s.d. (in c,e-g ). P values calculated using one-way or two-way ANOVA for multiple comparisons or two-tailed Student’s t -tests for unpaired comparisons.

Article Snippet: For measurement of IL-1β levels, monocytes were additionally treated with ATP (5 mM) for 45 min following LPS stimulation, pelleted at 500 g for 5 min, and supernatant IL-1β levels assayed using human IL-1β DuoSet ELISA kit (R&D Systems, #DY201).

Techniques: Phospho-proteomics, Western Blot, Incubation, Concentration Assay, Activity Assay, Quantitative Proteomics, Two Tailed Test

Journal: Nature chemical biology

Article Title: A druggable redox switch on SHP1 controls macrophage inflammation

doi: 10.1038/s41589-026-02163-8

Figure Lengend Snippet: a , Gating strategy for flow cytometry analysis of healthy donors or RA and MS patients PBMCs isolated monocytes using CD45, CD14 and CD16 staining ( n = 3 for each healthy and patient group). b , Percentage of CD45+CD14+CD16- monocyte population of interest ( n = 3 for each healthy and patient group). c , Pro-inflammatory cytokine TNF, IL-6 and IL-1β levels in cell supernatants of healthy donors and RA and MS patients monocytes pre-incubated 3 h with SCA1, SCA9, SCA7 and SCA25 (20 μM) followed by 6 h LPS stimulation (100 ng/ml) ( n = 3 for each healthy and patient group). For measurement of pro-inflammatory cytokine IL-1β levels in cell supernatants, cells were additionally treated with adenosine triphosphate (ATP, 5 mM) for 45 min following LPS treatment. Data are mean ± s.e.m. P values calculated using two-tailed Student’s t -test for unpaired comparison or two-way ANOVA for multiple comparisons.

Article Snippet: For measurement of IL-1β levels, monocytes were additionally treated with ATP (5 mM) for 45 min following LPS stimulation, pelleted at 500 g for 5 min, and supernatant IL-1β levels assayed using human IL-1β DuoSet ELISA kit (R&D Systems, #DY201).

Techniques: Flow Cytometry, Isolation, Staining, Incubation, Two Tailed Test, Comparison