Mouse IL-33 Antibody Best Seller

Detection of IL‑33 in Mouse Splenocytes by Flow Cytometry.

Mouse splenocytes were treated for 5 hours with 50 ng/mL PMA and 1 µg/mL Ca²⁺ionomycin then stained with Goat Anti-Mouse IL-33 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3626, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Proliferation Induced by IL‑33 and Neutralization by Human IL‑33 Antibody.

Recombinant Mouse IL-33 (Catalog # 3626-ML) stimulates proliferation in the D10.G4.1 mouse helper T cell line in a dose-dependent manner (orange line), as measured by Resazurin (Catalog # AR002). Proliferation elicited by Recombinant Mouse IL-33 (0.25 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse IL-33 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3626). The ND₅₀ is typically 10-50 ng/mL.

Detection of Mouse IL-33 by Immunohistochemistry

IL-33/ST2 axis in tendon healing in vivo.Kinetics of (a) Il33 and (b) soluble St2 gene expression post injury. Data are mean fold change±s.d. showing relative expression to 18s housekeeping gene (pooled data from four mice per group performed on four sequential occasions, n=16 per condition). *P<0.05, **P<0.01 versus control (WT) mice (ANOVA) +P<0.05,++P<0.01, NS not significant; control (WT) injured versus St2−/− injured. (c) Immunohistochemistry showing IL-33 and ST2 expression in tendon biopsies of WT and St2−/− mice day 1 post injury. IgG control shown in bottom left of pictures. Black horizontal line indicates 50 μm. (d) Col3 mRNA and (e) Collagen 3 protein levels in tendon biopsies of WT and St2−/− mice post injury. (f) Col1 mRNA and (g) Collagen 1 protein levels in tendon biopsies of WT and St2−/− mice post injury. For (d–g) data are mean±s.d. of duplicate samples, representative of four mice per condition (n=16). *P<0.05, **P<0.01 versus control (WT) mice. +P<0.05, ++P<0.01 control (WT) injured versus St2−/− injured mice (ANOVA). mRNA graphs show relative expression to 18s housekeeping gene. (h) Percentage change in tendon strength for WT and St2−/− mice post injury. Data are mean±s.d., representative of four mice per condition (n=16). *P<0.05, **P<0.01 versus control mice. #P<0.05 St2−/− injured versus WT injured mice (Mann–Whitney U-test). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25857925), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse IL-33 by Western Blot

HMGB1 increases IL-33 expression and acts as an autocrine factor in enhancing IL-33 expression in a partial TLR-4 dependent fashion during cyclic stretch.(A) MLE-12 cells were treated with 3 μg/ml HMGB1 or control solution before 6h cyclic stretch and IL-33 production was measured by ELISA. (B) MLE-12 cells were treated with 10 μg/ml HMGB1 neutralizing antibody (2G7) or control solution before 6h cyclic stretch and IL-33 production was detected by Western blot. beta-actin served as loading control. (C) MLE-12 cells transfecting with non-specific control siRNA or TLR-4-specific siRNA were treated with 3 μg/ml HMGB1 or control solution before stretch. IL-33 production in each group was measured with ELISA. **P<0.01, ***P<0.001 when compared between groups denoted by horizontal lines. Each stretching group collected at least from three wells and represented a single experiment, the bar graphs illustrate data representative of three independent experiments. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28898270), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human IL-33 by Immunohistochemistry

IL-33/ST2 expression in human tendon.(a) IL-33, (b) soluble ST2 (sST2) and (c) mST2 mRNA expression in human tendon samples. Fold change in gene expression of IL-33, sST2 and mST2 in control (semi-membranosus tendon, n=10), torn supraspinatus tendon (established pathology) and matched subscapularis human tendon samples (early pathology; n=17). Data are mean±s.d. relative to the housekeeping gene18S (mean of duplicate analysis). *P<0.05, **P<0.01, ***P<0.001 versus control (Student's t-test). (d) Immunostaining of IL-33 and ST2 in control (n=10), torn tendon (n=17) and early tendinopathy (n=17). Graphs illustrate modified Bonar scoring based on 10 high-power fields. Data are mean±s.d. *P<0.05, **P<0.01 versus control (Student's t-test). Scale bar, 65 μm. (e) Fold change in gene expression of IL33 and ST2 24 h post incubation with tumour necrosis factor (TNF-alpha), IL-1 beta or in combination depicting relative expression to media alone utilizing housekeeping gene GAPDH. Data are mean±s.d. of triplicate samples, representative of three individual patient samples. *P<0.05, **P<0.01 versus control (media) (Student's t-test). (f) IL-33 immunostaining of human tendon explants cultured for 24 h with medium (control), 100 ng ml−1 TNF alpha or 100 ng ml−1 TNF alpha+100 ng ml−1 IL-1 beta. (g) Fold change in COL1 and COL3 mRNA expression in human tendon explants cultured for 24 h with rhIL-33, relative to housekeeping gene GAPDH. (h) Time course of COL1 and COL3 mRNA expression following incubation with rhIL-33, relative to housekeeping gene GAPDH. (i) Collagen 1 and 3 protein expression in human tendon explants 24 h post incubation with rhIL-33. For (g–i) data are mean±s.d. of triplicate samples, representative of three individual patients. *P<0.05, **P<0.01 versus control (Student's t-test). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25857925), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Mouse IL-33 Antibody by Immunohistochemistry

Prior Nb exposure results in earlier and exacerbated HSV-2 pathology. (I and J) (I) Vaginal IL-33 measured by ELISA (day 2) and (J) IF staining of vaginal tissue (day 3; n = 4). Data are representative of two independent experiments with 4–6 mice per group (mean ± SEM). Statistical significance was calculated by two-way analysis of variance (ANOVA) with Bonferroni correction for multiple comparisons and Mann-Whitney t test. ∗p ≤ 0.05, ∗∗p ≤ 0.01, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.001, ns, not significant. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33857419), licensed under a CC-BY license. Not internally tested by R&D Systems.