primary antibody (mouse anti-osteocalcin)  (Thermo Fisher)

Journal: Nature Communications

Article Title: High-throughput bioprinting of spheroids for scalable tissue fabrication

doi: 10.1038/s41467-024-54504-7

Figure Lengend Snippet: A HITS-Bio setup under surgical settings. Inset images (1–2) demonstrate deposited BONink and spheroid placement, (ii) created calvarial defects ( ~ 5 mm in a diameter), and (iii) bioprinted bone constructs with BONink and spheroids. Scale bar: 5 mm. B Visualization of newly regenerated bone in calvarial defects with transverse and sagittal planes at Weeks 3 and 6 via µCT, including empty, BONink only, low-density and high-density groups. Scale bar: 1 mm. C Relevant quantification for bone regeneration within the defect examined at Weeks 3 and 6 ( n = 7 independent defects, one-way ANOVA, all other shown comparisons p < 0.0001), including (i) BV/TV (new bone volume to total bone volume, %) (Week 3: a vs. b p = 0.35139, a vs. c p = 0.00074, a vs. d p = 0.00545, b vs. c p = 0.04183, b vs. d p = 0.20350, c vs. d p = 0.85065, Week 6: a vs. b p = 0.60072, a vs. c p = 0.00186, a vs. d p = 0.00212, b vs. c p = 0.03614, b vs. d p = 0.04048, c vs. d p = 0.99995), (ii) normalized BMD (bone mineral density, %) (Week 3: a vs. b p = 0.045, b vs. c p = 0.03277, b vs. d p = 0.07177, c vs. d p = 0.98269, Week 6: a vs. b p = 0.63421, a vs. c p = 0.00473, a vs. d p = 0.00528, b vs. c p = 0.07204, b vs. d p = 0.07910, c vs. d p = 0.99996), (iii) bone coverage area (%) (Week 3: a vs. b p = 0.06611, b vs. c p = 0.00656, b vs. d p = 0.00464, c vs. d p = 0.99893, Week 6: a vs. b p = 0.11033, b vs. c p = 0.00259, b vs. d p = 0.00032, c vs. d p = 0.83536) (The outlier was not included in the data plot), and (iv) scores for bony bridging (Week 3: a vs. b p = 0.03791, b vs. c p = 0.01013, b vs. d p = 0.01013, c vs. d p = 1.0, Week 6: a vs. b p = 0.04481, b vs. c p = 0.00326, b vs. d p = 0.00326, c vs. d p = 1.0). D Mechanical properties of the retrieved defect area 6 weeks after the surgery ( n = 3, 4, 4, 3 independent defects (from left to right), one-way ANOVA) (Shear yield strength: a vs. b p = 0.97205, a vs. c p = 0.95577, a vs. d p = 0.14142, b vs. c p = 0.74719, b vs. d p = 0.05602, c vs. d p = 0.23294) (Modulus of resilience: a vs. b p = 0.98816, a vs. c p = 0.99588, a vs. d p = 0.00672, b vs. c p = 0.99957, b vs. d p = 0.00692, c vs. d p = 0.00598) (Shear modulus: a vs. b p = 0.65248, a vs. c p = 0.95136, a vs. d p = 0.94709, b vs. c p = 0.31149, b vs. d p = 0.92926, c vs. d p = 0.69672). E Histomorphometric characterization of sectioned defect after decalcification and stained for H&E (scale bar: 500 µm), MT (scale bar: 1 mm), and IHC (P1NP and OCN) (scale bar: 500 µm). Representative images were obtained from at least three independent repetitions. Data are presented as mean ± SD where * p < 0.05, ** p < 0.01, and *** p < 0.001. Source data are provided as a Source Data file.

Article Snippet: To detect the bone tissue, sections were incubated with a mouse anti-procollagen 1 N-Peptide (P1NP) primary antibody (1:250 in 2.5% NGS, MA5-51183, Invitrogen), mouse anti- osteocalcin (OCN) primary antibody (1:200 in 2.5% NGS, 33-5400, Invitrogen), mouse anti-RUNX2 (runt-related transcription factor 2) primary antibody (1:20 in 2.5% NGS, ab76956; Abcam) and a rabbit anti-Sp7/Osterix (OSTERIX) primary antibody (1:100 in 2.5% NGS; ab209484, Abcam) or a rabbit anti-bone sialoprotein (BSP) primary antibody (1:100 in 2.5% NGS; ab52128, Abcam) overnight.

Techniques: Construct, Shear, Staining

Journal: Bone reports

Article Title: Role of cartilage and bone matrix regulation in early equine osteochondrosis.

doi: 10.1016/j.bonr.2023.101653

Figure Lengend Snippet: Fig. 5. Photomicrographs depicting protein expression for osteocalcin following immunohistochemical localization. A) Osteochondral sample of foal with confirmed OC showing moderate to strong osteocalcin expression in chondrocytes along the osteochondral junction and in within the deeper cartilage layers. B) Negative control for (A) following substitution of mouse monoclonal osteocalcin antibody with nonimmune serum. C) Osteochondral samples of normal foal showing similar results (bar = 100 μm).

Article Snippet: Immunohistochemistry (IHC) was performed on paraffin-embedded osteochondral sections using mouse monoclonal primary antibodies for Osteocalcin (cat#MAB1419, R&D Systems, Inc., Minneapolis, MN) and Collagen type IIB (clone M2139, Thermo Fisher Scientific, Waltham, MA) and rabbit polyclonal primary antibodies for Lubricin (ab28484, Abcam, Cambridge, UK) and SOX9 (PA5-23383, Thermo Fisher Scientific, Waltham, MA).

Techniques: Expressing, Immunohistochemical staining, Negative Control

Journal: Biomaterials

Article Title: The use of heparin/polycation coacervate sustain release system to compare the bone regenerative potentials of 5 BMPs using a critical sized calvarial bone defect model

doi: 10.1016/j.biomaterials.2022.121708

Figure Lengend Snippet: Effects of 5 different BMPs on the osteogenic differentiation of hMDSCs at day 28 (3D pellet culture). A. MicroCT 3D images showed that the mineralized pellets in 5 BMPs groups were larger than the CTL group. Scale bar=1mm. B. Quantification of mineralized pellet volume. All BMPs groups showed higher mineralized pellet volume compared to the CTL group. The mineralized pellet volume of the BMP2 group was significantly higher than the BMP6 and BMP7 groups. *P<0.05, ** P<0.01, ****, P<0.0001. C. Mineralized pellet density quantification. No statistical difference between BMP groups and the CTL group was found. D. Von Kossa staining. Brown-black color indicates mineralization. All pellets have regions peeled off due to high mineralization in the white region of pellets. E. Immunohistochemistry of osteocalcin for osteogenic pellets. Brown color indicates osteocalcin expression. The mineralized parts of the pellets demonstrate crystal purple color intermingled with osteocalcin brown staining. Scale bars=100μm.

Article Snippet: Osteocalcin immunohistochemistry using a mouse anti-human osteocalcin primary antibody (MAB1419, 1:100, R&D Systems) was also performed, as previously described [ 27 ].

Techniques: Staining, Immunohistochemistry, Expressing