mouse anti il 1r8 antibody  (Santa Cruz Biotechnology)

Journal: International Journal of Molecular Medicine

Article Title: IL-37/IL-1R8 blocks keratinocyte acantholysis via suppressing ADAM17/EGFR

doi: 10.3892/ijmm.2026.5793

Figure Lengend Snippet: IL-37/IL-1R8 protects HaCaT cells from keratinocyte dissociation and apoptosis through the ADAM17/EGFR pathway. (A) The interaction between IL-37 and IL-1R8 was analyzed using co-immunoprecipitation. (B) HaCaT cells were transfected with IL-1R8 siRNA and the transfection efficiency were detected by western blotting. (C) HaCaT cells were transfected with IL-18Rα siRNA and the transfection efficiency were detected by western blotting. (D) IL-1R8 siRNA or IL-18Rα transfected HaCaT were treated with anti-Dsg-3 and IL-37 recombinant protein. The number of fragments was analyzed by cell dissociation assay. (E and F) Cell apoptosis was analyzed by flow cytometry. (G) Protein expression levels of Bcl-2 and Bax were determined by western blotting. & P<0.05 vs. si-NC; * P<0.05 vs. anti-Dsg3 + IL-37 + si-NC group. Data are presented as mean ± SD, n=3 biological independent replicates. One-way ANOVA with Bonferroni's post-hoc test was used for multiple group comparisons. IL, interleukin; ADAM17, TNF-alpha-converting enzyme; EGFR, epidermal growth factor receptor; si, short interfering.

Article Snippet: Following this, the membranes were incubated with the following specific primary antibodies overnight at 4°C: Rabbit anti-IL-37 monoclonal antibody (cat. no. ab278499; Abcam), rabbit anti-Bax monoclonal antibody (cat. no. ab32503; Abcam), rabbit anti-Bcl-2 polyclonal antibody (cat. no. ab59348; Abcam), rabbit anti-caspase 3 antibody (cat. no. 9662; Cell Signaling Technology, Inc.), rabbit anti-cleaved caspase3 antibody (cat. no. 9661; Cell Signaling Technology, Inc.), mouse anti-IL-1R8 antibody (cat. no. sc-271864; Santa Cruz Biotechnology, Inc.), mouse anti-IL-18Rα antibody (cat. no. PA5-115404; Thermo Fisher Scientific, Inc.), rabbit anti-EGFR phospho Y1173 antibody (cat. no. ab5652; Abcam), rabbit anti-EGFR antibody (cat. no. ab52894; Abcam), rabbit anti-ADAM17 polyclonal antibody (cat. no. ab39162; Abcam), anti-phosphorylated (p-) ERK1/2 antibody (cat. no. ab201015; Abcam), anti-ERK1/2 antibody (cat. no. ab184699; Abcam), anti-p-AKT (Ser473) antibody (cat. no. 9271; Cell Signaling Technology, Inc.), anti-AKT antibody (cat. no. 4691; Cell Signaling Technology, Inc.), anti-p-STAT3 antibody (cat. no. ab76315; Abcam), and anti-STAT3 antibody (cat. no. ab68153; Abcam).

Techniques: Immunoprecipitation, Transfection, Western Blot, Recombinant, Flow Cytometry, Expressing

Journal: International Journal of Molecular Medicine

Article Title: IL-37/IL-1R8 blocks keratinocyte acantholysis via suppressing ADAM17/EGFR

doi: 10.3892/ijmm.2026.5793

Figure Lengend Snippet: IL-37/IL-1R8 protects HaCaT cells from keratinocyte dissociation and apoptosis through the ADAM17/EGFR pathway. (A) Protein expression levels of ADAM17, p-EGFR, total EGFR as well as EGFR downstream protein p-AKT, p-ERK1/2 and p-STAT3 levels were determined by western blotting. HaCaT cells were treated with EGFR activator NSC228155. (B) Cell dissociation was analyzed by cell dissociation assay. (C) Cell apoptosis was analyzed by flow cytometry. (D) Protein expression levels of Bcl-2 and Bax were analyzed by western blotting. (E) HaCaT was transfected with ADAM17 siRNA and the transfection efficiency was detected by western blotting. HaCaT cells were transfected with ADAM17 siRNA or treated with 1 μ M of EGFR inhibitor AG1478 for 30 min followed by treated with AK23. (F) Cell dissociation was analyzed by cell dissociation assay. (G) Cell apoptosis was analyzed by flow cytometry. # P<0.05 vs. Control group; & P<0.05 vs. anti-Dsg3 group; @ P<0.05 vs. anti- Dsg3 + IL-37 + si-NC group; * P<0.05; Data are presented as mean ± SD, n=3 biological indepen- dent replicates. One-way ANOVA with Bonferroni's post-hoc test was used for multiple group comparisons. IL, interleukin; ADAM17, TNF-alpha-converting enzyme; EGFR, epidermal growth factor receptor; p- phosphorylated; STAT, signal transducer and activator of transcription; si, short interfering.

Article Snippet: Following this, the membranes were incubated with the following specific primary antibodies overnight at 4°C: Rabbit anti-IL-37 monoclonal antibody (cat. no. ab278499; Abcam), rabbit anti-Bax monoclonal antibody (cat. no. ab32503; Abcam), rabbit anti-Bcl-2 polyclonal antibody (cat. no. ab59348; Abcam), rabbit anti-caspase 3 antibody (cat. no. 9662; Cell Signaling Technology, Inc.), rabbit anti-cleaved caspase3 antibody (cat. no. 9661; Cell Signaling Technology, Inc.), mouse anti-IL-1R8 antibody (cat. no. sc-271864; Santa Cruz Biotechnology, Inc.), mouse anti-IL-18Rα antibody (cat. no. PA5-115404; Thermo Fisher Scientific, Inc.), rabbit anti-EGFR phospho Y1173 antibody (cat. no. ab5652; Abcam), rabbit anti-EGFR antibody (cat. no. ab52894; Abcam), rabbit anti-ADAM17 polyclonal antibody (cat. no. ab39162; Abcam), anti-phosphorylated (p-) ERK1/2 antibody (cat. no. ab201015; Abcam), anti-ERK1/2 antibody (cat. no. ab184699; Abcam), anti-p-AKT (Ser473) antibody (cat. no. 9271; Cell Signaling Technology, Inc.), anti-AKT antibody (cat. no. 4691; Cell Signaling Technology, Inc.), anti-p-STAT3 antibody (cat. no. ab76315; Abcam), and anti-STAT3 antibody (cat. no. ab68153; Abcam).

Techniques: Expressing, Western Blot, Flow Cytometry, Transfection, Control

Journal: Arthritis Research & Therapy

Article Title: Interleukin 37 limits monosodium urate crystal-induced innate immune responses in human and murine models of gout

doi: 10.1186/s13075-016-1167-y

Figure Lengend Snippet: Contribution of the Mertk inhibitor to the IL-37-mediated anti-inflammatory effect in monosodium urate ( MSU )-induced models in vitro and in vivo. a – c Concentration of secreted IL-1β, IL-8 and CCL2 in THP-1 macrophages treated with or without recombinant human IL-37 (rhIL-37) for 3 h, followed by incubation for 1 h with or without Mertk inhibitor and then incubated with lipopolysaccharide (LPS) or MSU separately for a further 18 h; * P < 0.05. d Different dosage of rhIL-37 was given preventively or therapeutically with or without Mertk inhibitor intervention in mice with gouty arthritis, and foot thickness was evaluated; * P < 0.05. e , f Histopathological analysis by H&E staining in a joint from the group treated with rhIL-37 treatment and Mertk inhibitor intervention (×100 original magnification ( e ) and × 200 original magnification ( f ); arrow inflammation in soft tissue and joint space. g – k The protein level of Smad3, IL-1R8, S​OCS3 and NLRP3 was verified by western blotting in the IL-37 treatment groups with or without Mertk inhibitor intervention. Protein levels in different groups were expressed as a ratio to that of corresponding glyceraldehyde-3-phosphate dehydrogenase ( GAPDH ); * P < 0.05 ** P < 0.01

Article Snippet: We used rabbit polyclonal anti-mouse NLRP3 antibody, anti-mouse Smad3, anti-mouse IL-1R8, anti-mouse S OCS3 (all antibodies were from Santa Cruz Biotechnology, Dallas, TX, USA) and rabbit polyclonal anti-mouse GAPDH antibody (Cell Signaling Technology, Danvers, MA, USA) as primary antibody and anti-rabbit IgG HRP-linked antibody (Cell Signaling Technology) as secondary antibody.

Techniques: In Vitro, In Vivo, Concentration Assay, Recombinant, Incubation, Staining, Western Blot