sclerostin antibody  (Novartis)

Journal: JBMR Plus

Article Title: The orally available SIK2/SIK3 inhibitor SK-124 increases bone mass in hypogonadal male mice

doi: 10.1093/jbmrpl/ziag032

Figure Lengend Snippet: Pharmacological inhibition of SIK2/SIK3 increases bone formation. (A) Representative H&E from tibia histologic sections from all treatment groups. Scale bar: 100 μm. (B) Representative fluorochrome-labeled femur histologic cross sections. Primary spongiosa (upper panel) and metaphyseal cortical (lower panel). Highly magnified double-calcein labeling images from all treatment groups. Scale bar: 100 μm. (C) Quantification of bone dynamic histomorphometry histological section derived measurements: MS/BS, MAR, and BFR were measured using the double calcein labels, which were injected at day 9 and day 2 before sacrifice timepoints. (D) Representative tibia sclerostin immunohistochemistry (DAB staining) sections from sham vehicle and all ORX treatment groups. Scale bar: 20 μm. (E) Quantification of Sost positive cells from sclerostin immunohistochemistry staining. Positive cells were counted at midshaft cortical region. (F) Relative gene expression levels of Sost to β- actin (left) and Tnfsf11 (right) to β- actin from marrow flushed cortical bone RNA from sham vehicle and all ORX treatment groups. All data were analyzed using one-way ANOVA followed by Dunnett’s post-hoc tests within sham or ORX groups, vs vehicle ( * p < .05; ** p < .01; *** p < .001). All values are indicated as mean ± SD. For MS/BS and endo-cortical MAR of panel C, n = 8 per group for vehicle and SK-124. n = 7 per group for PTH. For MAR and BFR of panel C, n = 8 per group for vehicle and SK-124. n = 7 per group for PTH in sham groups. n = 8 per group for ORX vehicle, n = 7 per group for ORX SK-124, and n = 4 per group for ORX PTH. n = 3 per group for panel E. For panel F, n = 5 per group for ORX PTH. n = 6 per group for ORX vehicle. N = 7 per group for sham vehicle and ORX SK-124.

Article Snippet: Next, slides were blocked in TNB buffer (Perkin-Elmer), stained with anti-sclerostin biotinylated antibody (1:50, R&D Systems, BAF1589) overnight at 4 °C.

Techniques: Inhibition, Labeling, Derivative Assay, Injection, Immunohistochemistry, Staining, Gene Expression

Journal: JBMR Plus

Article Title: Generation and characterization of a novel inducible Sost_P2A_CreERT2 mouse model with high specificity for osteocytes

doi: 10.1093/jbmrpl/ziag010

Figure Lengend Snippet: Insertion of the Sost_P2A_CreERT2 transgene does not cause a bone phenotype in heterozygous mice but results in increased bone density in homozygous mice. (A) The insertion site of the Sost_P2A_CreERT2 transgene sequence. (B) Sost mRNA expression is significantly downregulated in the long bones of homozygous Sost_P2A_CreERT2 mice but not heterozygous mice. (C) Whole body BMC and BMD are significantly increased in the homozygous but not heterozygous Sost_P2A_CreERT2 mice. **** p = <.0001, *** p = <.001, ** p = <.01, * p = <.05, one-way ANOVA with Tukey’s post hoc test.

Article Snippet: Sclerostin antibody (AF1589, R&D Systems) was diluted 1:200 in Primary Ab buffer (10 mL 1× PBS, 100 mg BSA, and 20 μL Triton-X).

Techniques: Sequencing, Expressing

Journal: JBMR Plus

Article Title: Generation and characterization of a novel inducible Sost_P2A_CreERT2 mouse model with high specificity for osteocytes

doi: 10.1093/jbmrpl/ziag010

Figure Lengend Snippet: Cre activation in the bones of male 5 mo Sost_P2A_CreERT mice following 5 doses of 75 mg/kg tamoxifen administration. (A) Low magnification image of the cortical bone at the midshaft of the femur (A) and trabecular bone at the distal femur (B) of a 5 mo Sost_P2A_CreERT2 +/− /Ai9 +/− mouse after tamoxifen injections. Higher magnification images of the femoral cortical bone (C), trabecular bone (D), vertebral trabecular bone (E), and calvaria (F) of tamoxifen and vehicle injected animals. (G) Brightfield and fluorescent images of selected soft tissues showing lack of TdT expression in the non-osseous tissues. Abbreviations: BM, bone marrow; CB, cortical bone; PB, parietal bone; PC, pericranial surface; TB, trabecular bone. Periosteal surfaces are delineated by a white dashed line and endosteal surfaces by a yellow dashed line. Scale bar = 200 μm.

Article Snippet: Sclerostin antibody (AF1589, R&D Systems) was diluted 1:200 in Primary Ab buffer (10 mL 1× PBS, 100 mg BSA, and 20 μL Triton-X).

Techniques: Activation Assay, Injection, Expressing

Journal: JBMR Plus

Article Title: Generation and characterization of a novel inducible Sost_P2A_CreERT2 mouse model with high specificity for osteocytes

doi: 10.1093/jbmrpl/ziag010

Figure Lengend Snippet: Cre activity is predominantly found in osteocytes closer to the periosteal surface compared to the endosteal surface. (A) Cortical bone from a male 2 mo Sost_P2A_CreERT2 +/− /Ai9 +/− mouse following 5 doses of 75 mg/kg tamoxifen administration. Scale bar = (50 μm). (B) Confocal microscopy image of osteocytes in cortical bone (100X) at the femoral mid-shaft of a male 5 mo Sost_P2A_CreERT2 +/− /Ai9 +/− mouse following 5 doses of 75 mg/kg tamoxifen. Periosteal surfaces are delineated by a white dashed line and endosteal surfaces by a yellow dashed line. Scale bar = 10 μm.

Article Snippet: Sclerostin antibody (AF1589, R&D Systems) was diluted 1:200 in Primary Ab buffer (10 mL 1× PBS, 100 mg BSA, and 20 μL Triton-X).

Techniques: Activity Assay, Confocal Microscopy

Journal: JBMR Plus

Article Title: Generation and characterization of a novel inducible Sost_P2A_CreERT2 mouse model with high specificity for osteocytes

doi: 10.1093/jbmrpl/ziag010

Figure Lengend Snippet: Cre activation in the bones of female 5 mo Sost_P2A_CreERT2 +/− /Ai9 +/− mice following 5 doses of 75 mg/kg tamoxifen administration. (A) Low magnification image of the cortical bone at the midshaft of the femur (A) and trabecular bone at the distal femur (B) of a 5 mo Sost_P2A_CreERT2 +/− /Ai9 +/− mouse after tamoxifen injections. Higher magnification images of the femoral cortical bone (C), trabecular bone (D), vertebral trabecular bone (E), and calvaria (F) of tamoxifen and vehicle injected animals. (G) Brightfield and fluorescent images of selected soft tissues showing lack of TdT expression in the non-osseous tissues. Abbreviations: BM, bone marrow; CB, cortical bone; PB, parietal bone; PC, pericranial surface; TB, trabecular bone. Periosteal surfaces are delineated by a white dashed line and endosteal surfaces by a yellow dashed line. Scale bar = 200 μm.

Article Snippet: Sclerostin antibody (AF1589, R&D Systems) was diluted 1:200 in Primary Ab buffer (10 mL 1× PBS, 100 mg BSA, and 20 μL Triton-X).

Techniques: Activation Assay, Injection, Expressing

Journal: JBMR Plus

Article Title: Generation and characterization of a novel inducible Sost_P2A_CreERT2 mouse model with high specificity for osteocytes

doi: 10.1093/jbmrpl/ziag010

Figure Lengend Snippet: Cre activity is induced in the ascending aorta of tamoxifen injected, but not vehicle injected Sost_P2A_CreERT2 +/− /Ai9 +/− mice. (A) Fluorescent and brightfield overlayed images of ascending and distal aorta from 5 mo male Sost_P2A_CreERT2 +/− /Ai9 +/− mice following 5 injections with 75 mg/kg tamoxifen or vehicle control. (B) Quantitation of TdT positive area in the ascending aorta in male and female Sost_P2A_CreERT2 +/− /Ai9 +/− . Scale bar = 200 μm. ** p = <.01, 2-tailed Student’s t -test.

Article Snippet: Sclerostin antibody (AF1589, R&D Systems) was diluted 1:200 in Primary Ab buffer (10 mL 1× PBS, 100 mg BSA, and 20 μL Triton-X).

Techniques: Activity Assay, Injection, Control, Quantitation Assay

Journal: JBMR Plus

Article Title: Generation and characterization of a novel inducible Sost_P2A_CreERT2 mouse model with high specificity for osteocytes

doi: 10.1093/jbmrpl/ziag010

Figure Lengend Snippet: Lowering the tamoxifen dose reduces Cre activity in the ascending aorta of Sost_P2A_CreERT2 +/− /Ai9 +/− . (A) Quantitation of TdT positive osteocytes in the bones from 2 mo Sost_P2A_CreERT2 +/− /Ai9 +/− mice injected with either 3 × 10 mg/kg or 5 × 75 mg/kg doses. The number of mice per group is shown in parentheses. TdT fluorescence (B) and quantitation of fluorescence area (C) in the ascending aorta of male Sost_P2A_CreERT2 +/− /Ai9 +/− mice. (D) TdT fluorescence and quantitation in the ascending aorta of female Sost_P2A_CreERT2 +/− /Ai9 +/− . Scale bar = 200 μm. *** p = <.001, 2-tailed Student’s t -test.

Article Snippet: Sclerostin antibody (AF1589, R&D Systems) was diluted 1:200 in Primary Ab buffer (10 mL 1× PBS, 100 mg BSA, and 20 μL Triton-X).

Techniques: Activity Assay, Quantitation Assay, Injection, Fluorescence

Journal: JBMR Plus

Article Title: Generation and characterization of a novel inducible Sost_P2A_CreERT2 mouse model with high specificity for osteocytes

doi: 10.1093/jbmrpl/ziag010

Figure Lengend Snippet: Further reducing the tamoxifen dosing does not affect Cre activity in the aorta of Sost_P2A_CreERT2 +/− /Ai9 +/− male mice. (A) Fluorescent images from the femur of 2 mo mice administered either 1, 2, or 3 injections with 10 mg/kg tamoxifen. Quantitation of TdT positive osteocytes in the cortical (B) and trabecular (C) bone. Fluorescent and brightfield overlayed images (D) and quantitation (E) of TdT expression in the ascending aorta in the 2 mo mice administered either 1, 2, or 3 injections with 10 mg/kg tamoxifen. Periosteal surfaces are delineated by a white dashed line and endosteal surfaces by a yellow dashed line. Scale bar = 200 μm.

Article Snippet: Sclerostin antibody (AF1589, R&D Systems) was diluted 1:200 in Primary Ab buffer (10 mL 1× PBS, 100 mg BSA, and 20 μL Triton-X).

Techniques: Activity Assay, Quantitation Assay, Expressing