◦c with anti osteocalcin ocn rabbit polyclonal antibody  (Servicebio Inc)

Journal: bioRxiv

Article Title: A Modular Bioinstructive Platform for Additive-Free, Topography-Driven Stem Cell Differentiation and Patterning

doi: 10.1101/2025.07.11.664383

Figure Lengend Snippet: A) Heatmap showing expression of selected genes associated with skeletogenesis across experimental conditions at days 3 and 14 post-seeding. B, C) Volcano plots displaying key differentially expressed genes associated with cytoskeleton, osteogenesis, and chondrogenesis in dimpled versus smooth microparticle cultures at day 3 (B) and day 14 (C) post-seeding (log 2 fold change > 1 and p adj < 0.05, N= 3 donors). D, E) Relative quantitative real-time PCR (qPCR) analysis of gene expression at specific time-points in serum-reduced medium, relative to 2D controls: RUNX2 (D; N= 5 donors) after 3 days, and SP7 (E; N= 5 donors) after 10 days. 2D positive control treated with 2 μM purmorphamine is shown as a reference point for RUNX2 expression. (F) Relative qPCR analysis of BGLAP expression after 10 days of either 300 nM KAAD-cyclopamine treatment or 0.06% ( v/v ) DMSO in serum-reduced medium, and relative to 2D vehicle-only controls (N= 5 donors). Statistical analysis was conducted using one-way ANOVA with Tukey’s multiple comparisons was applied. Values are presented as mean ± SD (** p < 0.01, *** p < 0.001, **** p < 0.0001). G, H) Representative maximum intensity projection confocal images of hMSCs stained for COL1A1 (green; G), and OCN (green; H) with nuclei counter-stained in blue (DAPI) after 10 days of culture (Scale bars: 50 μm). A single optical slice is used to represent brightfield for clarity. Abbreviations: SOX9, SRY-box transcription factor 9; COL2A1, collagen type II alpha 1 chain; COL10A1, collagen type x alpha 1 chain; COL1A1, collagen type I alpha 1; COL6A1, collagen type VI alpha 1 chain; ACAN, aggrecan; RUNX2, runt-related transcription factor 2; SP7, specificity protein 7, also known as osterix; MSX2, Msh homeobox 2; SPP1, secreted phosphoprotein 1, also known as osteopontin; POSTN, periostin; PAX9, paired box 9; BGLAP, bone gamma-carboxyglutamate protein, also known as osteocalcin; ALPL, alkaline phosphatase; TNC, tenascin-c; SCUBE3, signal peptide-CUB-EGF domain-containing protein 3 ; SERPINE2, serine protease inhibitor 2; GALNT1, N-acetylgalactosaminyltransferase 1; LEPR, leptin receptor; SOSTDC1, sclerostin domain-containing protein 1; CNMD , chondromodulin; BMP, bone morphogenetic protein; IGF-II, insulin-like growth factor 2; IGFBP, insulin-like growth factor binding protein; DAPI, 4′,6-Diamidino-2-phenylindole; OI media, Osteoinductive media; Pur, Purmorphamine; DIMP, Dimpled; SM, Smooth; DIMP_KD, Dimpled cultures treated with KAAD-cyclopamine .

Article Snippet: The following primary antibodies were used, goat anti-GLI-1 affinity purified polyclonal antibody (1:100; AF3455, R&D systems, RRID: AB_2247710), rabbit anti-osteocalcin (OCN) polyclonal antibody (1:75; AB10911, Millipore, RRID: AB_1587337), goat anti-type I collagen polyclonal antibody (1:200; #1310-01, SouthernBiotech, RRID: AB_2753206) and monoclonal anti-Vinculin (1:70; V4505, Sigma-Aldrich, RRID: AB_477617).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Gene Expression, Positive Control, Staining, Protease Inhibitor, Binding Assay

Journal: Bioengineering

Article Title: Comparison of Bioengineered Scaffolds for the Induction of Osteochondrogenic Differentiation of Human Adipose-Derived Stem Cells

doi: 10.3390/bioengineering11090920

Figure Lengend Snippet: hASC survival and differentiation in GelMA hydrogels. ( A ) CCK8 assay of hASCs encapsulated in the GelMA hydrogel at different time points (1 week intervals). Results are reported as mean ± SEM of n = 3 samples/group. ( B ) Representative confocal imaging of Dead/live fluorescence staining of hASCs in the GelMA hydrogel after 3 weeks of culture. Red cells are dead while green cells are alive. ( C ) Alcian blue staining of GelMA hydrogel after 24 h and 3 weeks of culture. ( D ) Alizarin red staining of GelMA hydrogel after 24 h and 3 weeks of culture. ( E , F ) Confocal imaging of immunofluorescence for COL2A1 (green) and OCN (red) in hASCs encapsulated in GelMA hydrogel after 3 weeks of culture.

Article Snippet: Briefly, samples were incubated with rabbit polyclonal anti-collagen type II alpha 1 (COL2A1) antibody (Merck) and rabbit polyclonal anti-osteocalcin (OCN) antibody (Thermo Fisher Scientific) for 1 h at room temperature.

Techniques: CCK-8 Assay, Imaging, Fluorescence, Staining, Immunofluorescence

Journal: Bioengineering

Article Title: Comparison of Bioengineered Scaffolds for the Induction of Osteochondrogenic Differentiation of Human Adipose-Derived Stem Cells

doi: 10.3390/bioengineering11090920

Figure Lengend Snippet: hASC survival and differentiation in PEGDA scaffold. ( A ) CCK8 assay of hASCs seeded in the PEGDA scaffold at different time points (1 week intervals). Results are reported as mean ± SEM of n = 3 samples/group. T test: * p < 0.05. ( B ) Representative confocal imaging of Dead/live fluorescence staining of hASCs seeded in the PEGDA scaffold after 3 weeks of culture. ( C ) Confocal imaging of immunofluorescence for COL2A1 (green) and OCN (red) in hASCs seeded in the PEGDA scaffold after 3 weeks of culture.

Article Snippet: Briefly, samples were incubated with rabbit polyclonal anti-collagen type II alpha 1 (COL2A1) antibody (Merck) and rabbit polyclonal anti-osteocalcin (OCN) antibody (Thermo Fisher Scientific) for 1 h at room temperature.

Techniques: CCK-8 Assay, Imaging, Fluorescence, Staining, Immunofluorescence

Journal: Bioengineering

Article Title: Comparison of Bioengineered Scaffolds for the Induction of Osteochondrogenic Differentiation of Human Adipose-Derived Stem Cells

doi: 10.3390/bioengineering11090920

Figure Lengend Snippet: hASC survival and differentiation in celery-based scaffold. ( A ) SEM imaging of a single hASC inside a niche of the celery-based scaffold. ( B ) CCK8 assay of hASCs seeded in the celery-based scaffold at different time points (1 week intervals). Results are reported as mean ± SEM of n = 3 samples/group. T test: * p < 0.05. ( C ) Representative confocal imaging of Dead/live fluorescence staining of hASCs seeded in the scaffold after 3 weeks of culture. ( D ) Confocal 3D stack and ( E ) the projection of hASC distribution inside the scaffold. ( F ) Confocal imaging of immunofluorescence for COL2A1 (green) and OCN (red) in hASCs seeded in the PEGDA scaffold after 3 weeks of culture.

Article Snippet: Briefly, samples were incubated with rabbit polyclonal anti-collagen type II alpha 1 (COL2A1) antibody (Merck) and rabbit polyclonal anti-osteocalcin (OCN) antibody (Thermo Fisher Scientific) for 1 h at room temperature.

Techniques: Imaging, CCK-8 Assay, Fluorescence, Staining, Immunofluorescence

Journal: PLOS ONE

Article Title: A comprehensive in vitro characterization of non-crosslinked, diverse tissue-derived collagen-based membranes intended for assisting bone regeneration

doi: 10.1371/journal.pone.0298280

Figure Lengend Snippet: Primer sequences.

Article Snippet: Primary rabbit polyclonal antibody anti-OCN (ab93876, Abcam, UK), mouse monoclonal antibody anti-Tubulin (sc-5286,Santa Cruz Biotechnology, USA), chemiluminescence suitable horseradish peroxidase-conjugated secondary antibodies (Santa Cruz Biotechnology, USA) and ECL system (Merck Millipore, Darmstadt Germany).

Techniques:

Journal: PLOS ONE

Article Title: A comprehensive in vitro characterization of non-crosslinked, diverse tissue-derived collagen-based membranes intended for assisting bone regeneration

doi: 10.1371/journal.pone.0298280

Figure Lengend Snippet: Evaluation of osteocalcin (OCN) protein level in bone isolated cells at t 0 (a) and after in vitro culture of cells on the membranes and on TCP (ctr) for 7 days (b). Results of the densitometric analysis: OCN protein expression normalized to tubulin expression for cells at t 0 and after 7days of culture on membranes 1–4 and on TCP (c). *p<0.05 vs t 0 .

Article Snippet: Primary rabbit polyclonal antibody anti-OCN (ab93876, Abcam, UK), mouse monoclonal antibody anti-Tubulin (sc-5286,Santa Cruz Biotechnology, USA), chemiluminescence suitable horseradish peroxidase-conjugated secondary antibodies (Santa Cruz Biotechnology, USA) and ECL system (Merck Millipore, Darmstadt Germany).

Techniques: Isolation, In Vitro, Expressing