primary antibodies to polyclonal goat anti-il-36α, anti-il-36β, anti-il-36γ, anti-il-36r, and anti-il-36ra  (Santa Cruz Biotechnology)

Journal: Communications Biology

Article Title: Bone mineral loss damages renal tubules in mice

doi: 10.1038/s42003-026-09603-0

Figure Lengend Snippet: Total RNA and formalin-fixed paraffin sections were prepared from the kidneys from mice used in Fig. . Relative mRNA levels of renal tubular damage markers were measured by quantitative RT-PCR. a Osteopontin. b Neutrophil gelatinase-associated lipocalin (Ngal). c Kidney injury molecule-1 (Kim-1). d Interleukin-36α (IL-36α). e Interleukin-6 (IL-6). Data are indicated as violin plots with the median and quartiles (dotted lines). N = 6 for each column. * P < 0.05, # P < 0.01 versus baseline by one-way ANOVA with Tukey’s multiple comparison test. f Plasma creatinine levels. Data are presented as means ± SD. N = 6 for each column. No statistical differences were observed between the groups by one-way ANOVA with Tukey’s multiple comparison test. g Representative kidney section images of immunohistochemistry using antibodies against osteopontin (upper panels) or IL-36α (lower panels) at the indicated time points after the last sRANKL or vehicle injection. Bar = 1 mm.

Article Snippet: After blocking, the sections were incubated with a goat polyclonal antibody against mouse IL-36α (diluted 1: 100 with PBS containing 1% BSA; AF2297; R&D Systems, Minneapolis, MN, USA), a mouse monoclonal antibody against mouse osteopontin (diluted 1: 100 with PBS containing 1% BSA; sc-21742; Santa Cruz, Dallas, TX, USA), or rat monoclonal antibody against mouse F4/80 (diluted 1: 100 with PBS containing 1% BSA; GTX26640; GeneTex, San Antonio, TX, USA) at room temperature for 1 h and then with Histofine Simple Stain Mouse MAX PO (Nichirei Corporation) at room temperature for 30 min. Immunoreactivity was visualized by incubation with the DAB substrate kit (Dako; Agilent Pathology Solutions).

Techniques: Quantitative RT-PCR, Comparison, Clinical Proteomics, Immunohistochemistry, Injection

Journal: Communications Biology

Article Title: Bone mineral loss damages renal tubules in mice

doi: 10.1038/s42003-026-09603-0

Figure Lengend Snippet: a The study design. Exogenous phosphate loading was induced by feeding a high phosphate diet (1.5% inorganic phosphate, “Pi”, HP diet) for 6 weeks, and endogenous phosphate loading was elicited by sRANKL administration using the triple dose regimen given every other week, respectively. b – f Relative mRNA levels of osteopontin ( OPN ), I L-36α, F4/80, Mcp-1 , and Collagen 1a1 quantified by qPCR. g Plasma FGF23 levels. Data are presented as means ± SD. N = 7–8 per group. Statistical significance was assessed by two-way ANOVA followed by Tukey’s multiple-comparison test; P valuses are indicated. Effect sizes of HP diet, sRANKL treatment, and their interaction were calculated from the ANOVA sums of squares and shown as partial η² (pη²).

Article Snippet: After blocking, the sections were incubated with a goat polyclonal antibody against mouse IL-36α (diluted 1: 100 with PBS containing 1% BSA; AF2297; R&D Systems, Minneapolis, MN, USA), a mouse monoclonal antibody against mouse osteopontin (diluted 1: 100 with PBS containing 1% BSA; sc-21742; Santa Cruz, Dallas, TX, USA), or rat monoclonal antibody against mouse F4/80 (diluted 1: 100 with PBS containing 1% BSA; GTX26640; GeneTex, San Antonio, TX, USA) at room temperature for 1 h and then with Histofine Simple Stain Mouse MAX PO (Nichirei Corporation) at room temperature for 30 min. Immunoreactivity was visualized by incubation with the DAB substrate kit (Dako; Agilent Pathology Solutions).

Techniques: Clinical Proteomics, Comparison

Journal: Communications Biology

Article Title: Bone mineral loss damages renal tubules in mice

doi: 10.1038/s42003-026-09603-0

Figure Lengend Snippet: Plasma levels of phosphate ( a ), calcium ( b ), calcium phosphate product ( c ), CPPs ( d ), FGF23 ( e ), and TRAcP-5b ( f ) of the mice from the normal gravity group (1 G) and the microgravity group (0 G). Relative mRNA levels of osteopontin ( g ) and interleukin-36α (IL-36α) ( h ) in the kidney. Data are presented as means ± SD. N = 6 for each column. P values by t test are indicated.

Article Snippet: After blocking, the sections were incubated with a goat polyclonal antibody against mouse IL-36α (diluted 1: 100 with PBS containing 1% BSA; AF2297; R&D Systems, Minneapolis, MN, USA), a mouse monoclonal antibody against mouse osteopontin (diluted 1: 100 with PBS containing 1% BSA; sc-21742; Santa Cruz, Dallas, TX, USA), or rat monoclonal antibody against mouse F4/80 (diluted 1: 100 with PBS containing 1% BSA; GTX26640; GeneTex, San Antonio, TX, USA) at room temperature for 1 h and then with Histofine Simple Stain Mouse MAX PO (Nichirei Corporation) at room temperature for 30 min. Immunoreactivity was visualized by incubation with the DAB substrate kit (Dako; Agilent Pathology Solutions).

Techniques: Clinical Proteomics

Journal: Communications Biology

Article Title: Bone mineral loss damages renal tubules in mice

doi: 10.1038/s42003-026-09603-0

Figure Lengend Snippet: Total RNA and formalin-fixed paraffin sections were prepared from the kidney samples from mice in Fig. . Relative mRNA levels of Osteopontin ( a ), Ngal , ( b ), Kim-1 ( c ), IL-36α ( d ), and matrix metalloprotease-3 ( MMP3 ) ( e ) were measured by quantitative RT-PCR. Data are indicated as violin plots with the median and quartiles (dotted lines). N = 7–12 for each column. P values by Kruskal–Wallis’s test with Dunn’s multiple-comparison test are indicated. f Representative kidney section images of hematoxylin-eosin staining (H&E) and immunohistochemistry using antibodies against osteopontin (OPN) or IL-36α from mice treated with sRANKL (upper panels) and mice co-treated with sRANKL and Risedronate (lower panels). Bar: 0.1 mm for H&E and 1 mm for immunohistochemistry.

Article Snippet: After blocking, the sections were incubated with a goat polyclonal antibody against mouse IL-36α (diluted 1: 100 with PBS containing 1% BSA; AF2297; R&D Systems, Minneapolis, MN, USA), a mouse monoclonal antibody against mouse osteopontin (diluted 1: 100 with PBS containing 1% BSA; sc-21742; Santa Cruz, Dallas, TX, USA), or rat monoclonal antibody against mouse F4/80 (diluted 1: 100 with PBS containing 1% BSA; GTX26640; GeneTex, San Antonio, TX, USA) at room temperature for 1 h and then with Histofine Simple Stain Mouse MAX PO (Nichirei Corporation) at room temperature for 30 min. Immunoreactivity was visualized by incubation with the DAB substrate kit (Dako; Agilent Pathology Solutions).

Techniques: Quantitative RT-PCR, Comparison, Staining, Immunohistochemistry