Human IL-1 beta/IL-1F2 Antibody

Detection of Human IL-1 beta/IL-1F2 by Simple Western^TM.

Simple Western lane view shows lysates of THP-1 human acute monocytic leukemia cell line untreated (-) or treated (+) with 200 nm PMA and 10 ug/ml LPS for 24 hrs and 3 hrs, respectively, loaded at 0.2 mg/mL. A specific band was detected for IL-1 beta/IL-1F2 at approximately 38 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human IL-1 beta/IL-1F2 Monoclonal Antibody (Catalog # MAB201) . This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Cell Proliferation Induced by IL‑1 beta/IL‑1F2 and Neutralization by Human IL‑1 beta/IL‑1F2 Antibody.

Recombinant Human IL-1 beta/IL-1F2 (Catalog # 201-LB) stimulates proliferation in the the D10.G4.1 mouse helper T cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human IL-1 beta/IL-1F2 (50 pg/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human IL-1 beta/IL-1F2 Monoclonal Antibody (Catalog # MAB201). The ND₅₀ is typically 0.001-0.003 µg/mL.

Detection of Human IL-1 beta/IL-1F2 by Western Blot

Hsp27 modulates IL-1 beta production in monocytes.(a) Immunoblot analysis of Hsp27 protein expression in classical (CL) and non-classical (NC) monocytes (left), and relative band densities from 2 combined experiments (right), with GAPDH as loading control. (b) Representative immunoblot of Hsp27 and pro-IL-1 beta (p31) in lysates from siRNA-transfected total monocytes for si-control (scrambled control siRNA) or si-Hsp27 (siRNA to Hsp27) either untreated or stimulated with LPS for 3 h. Data in (a) and (b) are cropped images and the corresponding full-length images are presentated in Supplementary Fig. 3. (c) Expression of IL-1 beta mRNA in siRNA-transfected total monocytes (n = 3). (d) Mature IL-1 beta release measured by ELISA of supernatants from siRNA-transfected total monocytes upon LPS stimulation in the presence or absence of BzATP (n = 4). (e) Fold change in IL-1 beta production by LPS and BzATP-treated total monocytes transfected with si-Hsp27, with respect to si-control. Data plotted are mean ± SEM, student t-test (a and b) and two-way ANOVA with Tukey’s multiple comparison test (c), *p < 0.05, **p < 0.01. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27976724), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human IL-1 beta/IL-1F2 by Western Blot

Non-classical monocytes secrete less mature IL-1 beta compared to other monocyte subsets following LPS and BzATP stimulation.(a) IL-1 beta ELISA on supernatants from monocyte subsets either untreated or stimulated with LPS for 24 h in the presence or absence of the P2X7 antagonist, A438079, followed by a secondary stimulation by BzATP in some conditions as indicated. Data plotted are mean ± SEM; n = 8. ****p < 0.0001 ***p < 0.001. ##p < 0.01 and #p < 0.05 is comparison to the respective classical and intermediate subsets treated with LPS + BzATP. Statistics are calculated based on Two-way ANOVA with Tukey’s multiple comparison test (*p < 0.05). (b) Representative cropped immunoblot images for mature 17 kDa (p17) IL-1 beta in culture supernatants of monocyte subsets stimulated as described in (a). The samples were derived from the same experiment and were ran on the same gel. Full-length images are shown in Supplementary Fig. 3. (c) YoPro-1 dye uptake was measured by flow cytometry to reflect level of P2X7R activity, and is shown as the fold change of BzATP-induced YoPro-1 uptake normalized to the respective untreated subset uptake. Data plotted are mean ± SEM; n = 6 based on one-way ANOVA with Tukey’s multiple comparison test. CL: classical, INT: intermediate, NC: non-classical. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27976724), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Rabbit IL-1 beta/IL-1F2 by Immunocytochemistry/Immunofluorescence

Immunohistochemistry for CD3, IL-1 beta and CD79 expression.Some expression for these markers were found in the CS and CSVT groups at 21 days (positive signal by brown staining). The same overall expression to the materials was seen at the other time points. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0136514), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IL-1 beta/IL-1F2 by Western Blot

Yptb effectors enable evasion of T3SS-dependent inflammasome activation in human cells. BMDMs (A), Caco-2 cells (B, D, and E), or primary hMDMs from three healthy human donors (C, F, and G) were infected with PBS (mock), WT Yptb, or delta6 Yptb. (A–C) Cell death was measured as percentage of cytotoxicity normalized to cells treated with 2% triton at 6 hpi. (D) Release of IL-18 into the supernatant was measured by ELISA at 6 hpi. (E) Lysates and supernatants collected 6 hpi were immunoblotted for IL-18 and beta-actin. (F) Release of IL-1 beta into the supernatant was measured by ELISA at 6 hpi. (G) Lysates and supernatants collected 6 hpi were immunoblotted for IL-1 beta and beta-actin. ns: not significant; *P < 0.05; **P < 0.01 by t-test (A–C) or one-way ANOVA (D and F). Shown are averages and error bars representing the standard deviation from at least three pooled experiments. ANOVA, analysis of variance; BMDMs, bone marrow-derived macrophages; ELISA, enzyme-linked immunosorbent assay; PBS, phosphate-buffered saline. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37615436), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IL-1 beta/IL-1F2 by Western Blot

Yptb effectors enable evasion of T3SS-dependent inflammasome activation in human cells. BMDMs (A), Caco-2 cells (B, D, and E), or primary hMDMs from three healthy human donors (C, F, and G) were infected with PBS (mock), WT Yptb, or delta6 Yptb. (A–C) Cell death was measured as percentage of cytotoxicity normalized to cells treated with 2% triton at 6 hpi. (D) Release of IL-18 into the supernatant was measured by ELISA at 6 hpi. (E) Lysates and supernatants collected 6 hpi were immunoblotted for IL-18 and beta-actin. (F) Release of IL-1 beta into the supernatant was measured by ELISA at 6 hpi. (G) Lysates and supernatants collected 6 hpi were immunoblotted for IL-1 beta and beta-actin. ns: not significant; *P < 0.05; **P < 0.01 by t-test (A–C) or one-way ANOVA (D and F). Shown are averages and error bars representing the standard deviation from at least three pooled experiments. ANOVA, analysis of variance; BMDMs, bone marrow-derived macrophages; ELISA, enzyme-linked immunosorbent assay; PBS, phosphate-buffered saline. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37615436), licensed under a CC-BY license. Not internally tested by R&D Systems.