primary antibody against ocn  (Santa Cruz Biotechnology)

Journal: GeroScience

Article Title: Age-associated methionine sulfoxide reductase A protects against valvular interstitial cell senescence and valvular calcification

doi: 10.1007/s11357-025-01675-w

Figure Lengend Snippet: MSRA expression is upregulated in CAVD. A H&E staining showing the normal and calcific human aortic valve tissues and immunohistochemical staining showing P21 and MSRA expression in the normal and calcific human aortic valve tissues ( n = 6 for each group), scale bar = 500 μm. B Western blot analysis and quantification of OCN, RUNX2, OPN, and MSRA protein expression in calcified VICs and controls ( n = 6 for each group). C Immunofluorescence staining and quantification of RUNX2 (red) and MSRA (green) in calcified VICs and controls ( n = 6 for each group), scale bar = 20 μm. D Flow cytometry analysis and the mean fluorescence intensity of RUNX2 and MSRA in calcified VICs and controls ( n = 4 for each group). Data are presented as mean ± SEM and compared by Student’s t -test. MSRA, methionine sulfoxide reductase A; CAVD, calcific aortic valve disease; H&E, hematoxylin-and eosin; OM, osteogenic medium; OCN, osteocalcin; RUNX2, runt-related transcription factor; OPN, osteopontin; VICs, valvular interstitial cells; DAPI, 4′,6-diamidino-2-phenylindole. NS, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against osteocalcin (OCN) (A20800, Abclonal, Wuhan, China, dilution 1:1000), runt-related transcription factor 2 (RUNX2) (A11753, Abclonal, Wuhan, China, dilution 1:1000), osteopontin (OPN) (ab214050, Abcam, Cambridge, USA, dilution 1:1000), cyclin-dependent kinase inhibitor 1 A (P21) (10,355–1-AP, Proteintech, Wuhan, China, dilution 1:1000), MSRA (14,547–1-AP, Proteintech, Wuhan, China, dilution 1:1000), TLR2 (66,645–1-Ig, Proteintech, Wuhan, China, dilution 1:1000), phosphorylated NF-κB p65 (3033, CST, Danvers, USA, dilution 1:1000), NF-κB p65 (8242, CST, Danvers, USA, dilution 1:1000), and GAPDH (A19056, Abclonal, Wuhan, China) were used.

Techniques: Expressing, Staining, Immunohistochemical staining, Western Blot, Immunofluorescence, Flow Cytometry, Fluorescence

Journal: GeroScience

Article Title: Age-associated methionine sulfoxide reductase A protects against valvular interstitial cell senescence and valvular calcification

doi: 10.1007/s11357-025-01675-w

Figure Lengend Snippet: MSRA alleviates osteogenic differentiation of human VICs in vitro. A Western blot analysis and quantification of OCN, RUNX2, OPN, and MSRA protein expression in VICs under osteogenic medium or control medium following MSRA silencing ( n = 6 for each group). B Immunofluorescence staining and quantification of RUNX2 (red) under osteogenic medium or control medium after MSRA silencing ( n = 6 for each group), scale bar = 20 μm. C Flow cytometry analysis and the mean fluorescence intensity of RUNX2 in VICs under osteogenic medium or control medium after MSRA silencing ( n = 4 for each group). D Western blot analysis and quantification of OCN, RUNX2, OPN, and MSRA protein expression in VICs under osteogenic medium or control medium following MSRA overexpression ( n = 6 for each group). E Immunofluorescence staining and quantification of RUNX2 (red) under osteogenic medium or control medium after MSRA overexpression ( n = 6 for each group), scale bar = 20 μm. F Flow cytometry analysis and the mean fluorescence intensity of RUNX2 in VICs under osteogenic medium or control medium after MSRA overexpression ( n = 4 for each group). Data are presented as means ± SEM and compared by Student’s t -test or one-way analysis of variance followed by Bonferroni post hoc test. MSRA, methionine sulfoxide reductase A; OM, osteogenic medium; OCN, osteocalcin; RUNX2, runt-related transcription factor; OPN, osteopontin; VICs, valvular interstitial cells; DAPI, 4′,6-diamidino-2-phenylindole. NS, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against osteocalcin (OCN) (A20800, Abclonal, Wuhan, China, dilution 1:1000), runt-related transcription factor 2 (RUNX2) (A11753, Abclonal, Wuhan, China, dilution 1:1000), osteopontin (OPN) (ab214050, Abcam, Cambridge, USA, dilution 1:1000), cyclin-dependent kinase inhibitor 1 A (P21) (10,355–1-AP, Proteintech, Wuhan, China, dilution 1:1000), MSRA (14,547–1-AP, Proteintech, Wuhan, China, dilution 1:1000), TLR2 (66,645–1-Ig, Proteintech, Wuhan, China, dilution 1:1000), phosphorylated NF-κB p65 (3033, CST, Danvers, USA, dilution 1:1000), NF-κB p65 (8242, CST, Danvers, USA, dilution 1:1000), and GAPDH (A19056, Abclonal, Wuhan, China) were used.

Techniques: In Vitro, Western Blot, Expressing, Control, Immunofluorescence, Staining, Flow Cytometry, Fluorescence, Over Expression

Journal: GeroScience

Article Title: Age-associated methionine sulfoxide reductase A protects against valvular interstitial cell senescence and valvular calcification

doi: 10.1007/s11357-025-01675-w

Figure Lengend Snippet: MSRA alleviates VIC senescence in vitro. A Western blot analysis and quantification of P21, OCN, RUNX2, OPN, and MSRA protein expression in VICs under H 2 O 2 stimulus after MSRA silencing ( n = 6 for each group). B Immunofluorescence staining and quantification of P21 (magenta) and RUNX2 (red) in VICs under H 2 O 2 stimulus after MSRA silencing ( n = 6 for each group), scale bar = 20 μm. C Western blot analysis and quantification of P21, OCN, RUNX2, OPN, and MSRA protein expression in VICs under H 2 O 2 stimulus after MSRA overexpression ( n = 6 for each group). D Immunofluorescence staining and quantification of P21 (magenta) and RUNX2 (red) in VICs under H 2 O 2 stimulus after MSRA overexpression ( n = 6 for each group), scale bar = 20 μm. Data are presented as means ± SEM and compared by Student’s t -test or one-way analysis of variance followed by Bonferroni post hoc test. MSRA, methionine sulfoxide reductase A; P21, cyclin-dependent kinase inhibitor 1 A; H 2 O 2 , hydrogen peroxide; OCN, osteocalcin; RUNX2, runt-related transcription factor; OPN, osteopontin; VICs, valvular interstitial cells; DAPI, 4′,6-diamidino-2-phenylindole. NS, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against osteocalcin (OCN) (A20800, Abclonal, Wuhan, China, dilution 1:1000), runt-related transcription factor 2 (RUNX2) (A11753, Abclonal, Wuhan, China, dilution 1:1000), osteopontin (OPN) (ab214050, Abcam, Cambridge, USA, dilution 1:1000), cyclin-dependent kinase inhibitor 1 A (P21) (10,355–1-AP, Proteintech, Wuhan, China, dilution 1:1000), MSRA (14,547–1-AP, Proteintech, Wuhan, China, dilution 1:1000), TLR2 (66,645–1-Ig, Proteintech, Wuhan, China, dilution 1:1000), phosphorylated NF-κB p65 (3033, CST, Danvers, USA, dilution 1:1000), NF-κB p65 (8242, CST, Danvers, USA, dilution 1:1000), and GAPDH (A19056, Abclonal, Wuhan, China) were used.

Techniques: In Vitro, Western Blot, Expressing, Immunofluorescence, Staining, Over Expression

Journal: GeroScience

Article Title: Age-associated methionine sulfoxide reductase A protects against valvular interstitial cell senescence and valvular calcification

doi: 10.1007/s11357-025-01675-w

Figure Lengend Snippet: MSRA alleviates osteogenic differentiation of VICs by inhibiting the TLR2/NF-κB pathway. A Western blot analysis and quantification of OCN, RUNX2, and OPN protein expression when VICs were cultured in osteogenic medium with or without TLR2 agonist Pam3CSK4 (0.1 μg/mL) after MSRA overexpression ( n = 6 for each group). B Immunofluorescence staining and quantification of RUNX2 (red) when human VICs were cultured in osteogenic medium with or without TLR2 agonist Pam3CSK4 after MSRA overexpression ( n = 6 for each group), scale bar = 20 μm. C Flow cytometry analysis and the mean fluorescence intensity of RUNX2 when human VICs were cultured in osteogenic medium with or without TLR2 agonist Pam3CSK4 after MSRA overexpression ( n = 4 for each group). D Alkaline phosphatase staining when VICs were cultured in osteogenic medium with or without TLR2 agonist Pam3CSK4 after MSRA overexpression ( n = 6 for each group), scale bar = 5 mm and 500 μm. Data are presented as means ± SEM and compared by Student’s t -test or one-way analysis of variance followed by Bonferroni post hoc test. MSRA, methionine sulfoxide reductase A; VICs, valvular interstitial cells; OM, osteogenic medium; OCN, osteocalcin; RUNX2, runt-related transcription factor; OPN, osteopontin; DAPI, 4′,6-diamidino-2-phenylindole. NS, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against osteocalcin (OCN) (A20800, Abclonal, Wuhan, China, dilution 1:1000), runt-related transcription factor 2 (RUNX2) (A11753, Abclonal, Wuhan, China, dilution 1:1000), osteopontin (OPN) (ab214050, Abcam, Cambridge, USA, dilution 1:1000), cyclin-dependent kinase inhibitor 1 A (P21) (10,355–1-AP, Proteintech, Wuhan, China, dilution 1:1000), MSRA (14,547–1-AP, Proteintech, Wuhan, China, dilution 1:1000), TLR2 (66,645–1-Ig, Proteintech, Wuhan, China, dilution 1:1000), phosphorylated NF-κB p65 (3033, CST, Danvers, USA, dilution 1:1000), NF-κB p65 (8242, CST, Danvers, USA, dilution 1:1000), and GAPDH (A19056, Abclonal, Wuhan, China) were used.

Techniques: Western Blot, Expressing, Cell Culture, Over Expression, Immunofluorescence, Staining, Flow Cytometry, Fluorescence

Journal: World Journal of Stem Cells

Article Title: EZH2, via an association with KDM2B, modulates osteogenic differentiation of root apical papillary stem cells

doi: 10.4252/wjsc.v17.i4.103482

Figure Lengend Snippet: Knockdown of enhancer of zeste homolog 2 inhibits steo/dentinogenic differentiation potential of human apical papillary stem cells. A: Quantitative polymerase chain reaction showed that the expression of enhancer of zeste homolog 2 (EZH2) was inhibited in human apical papillary stem cells (hSCAPs); B: Western blot analysis confirmed the knockdown of EZH2 in hSCAPs; C: Knockdown of EZH2 decreased alkaline phosphatase activity in hSCAPs; D and E: Alizarin red staining and quantitative calcium analysis demonstrated that knockdown of EZH2 inhibited mineralization in hSCAPs; F-H: Quantitative polymerase chain reaction showed that knockdown of EZH2 downregulated mRNA expression levels of bone sialoprotein (F), dentin sialophosphoprotein (G), and osteocalcin (H) in hSCAPs. GAPDH and ACTB was used as the internal controls. Data are presented as the mean ± SD ( n = 3). Statistical analysis was performed using Student’s t -test. a P ≤ 0.05, b P ≤ 0.01, c P ≤ 0.001. EZH2: Enhancer of zeste homolog 2; BSP: Bone sialoprotein; DSPP: Dentin sialophosphoprotein; OCN: Osteocalcin.

Article Snippet: Immunohistochemical analysis followed established protocols[ ] using primary antibodies against osteocalcin (OCN) (Bioss bs-4917R) and DSPP (Bioss bs10316R).

Techniques: Knockdown, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Activity Assay, Staining

Journal: World Journal of Stem Cells

Article Title: EZH2, via an association with KDM2B, modulates osteogenic differentiation of root apical papillary stem cells

doi: 10.4252/wjsc.v17.i4.103482

Figure Lengend Snippet: Overexpression of enhancer of zeste homolog 2 enhances osteo/dentinogenic differentiation potential of human apical papillary stem cells. A: Quantitative polymerase chain reaction showed that enhancer of zeste homolog 2 (EZH2) was overexpressed in human apical papillary stem cells (hSCAPs); B: Western blot analysis confirmed overexpression of EZH2 in hSCAPs; C: Overexpression of EZH2 increased alkaline phosphatase activity in hSCAPs; D and E: Alizarin red staining and quantitative calcium analysis results demonstrated that overexpression of EZH2 enhanced mineralization in hSCAPs; F-H: Quantitative polymerase chain reaction showed that overexpression of EZH2 upregulated mRNA expression levels of bone sialoprotein (F), dentin sialophosphoprotein (G), and osteocalcin (H) in hSCAPs; I: Hematoxylin-eosin staining and quantitative measurement showed that overexpression of EZH2 promoted bone/dentin-like tissue formation. Scale bar = 100 μm (B: Bone/dentin-like tissues; HA: Hydroxyapatite tricalcium carrier; CT: Connective tissue); J: Immunohistochemical staining and quantitative analysis of dentin sialophosphoprotein and bone sialoprotein. GAPDH and ACTB were used as the internal controls. Data are presented as the mean ± SD ( n = 3). Statistical analysis was performed using Student’s t -test. a P ≤ 0.05, b P ≤ 0.01, c P ≤ 0.001. EZH2: Enhancer of zeste homolog 2; BSP: Bone sialoprotein; DSPP: Dentin sialophosphoprotein; OCN: Osteocalcin.

Article Snippet: Immunohistochemical analysis followed established protocols[ ] using primary antibodies against osteocalcin (OCN) (Bioss bs-4917R) and DSPP (Bioss bs10316R).

Techniques: Over Expression, Real-time Polymerase Chain Reaction, Western Blot, Activity Assay, Staining, Expressing, Immunohistochemical staining

Journal: Biomaterials Translational

Article Title: Ångstrom-scale silver particle-infused hydrogels eliminate orthopedic implant infections and support fracture healing

doi: 10.12336/biomatertransl.2025.01.007

Figure Lengend Snippet: Abbreviations: AgÅPs: Ångstrom-scale silver particles; OCN: Osteocalcin; TRAP: Tartrate resistant acid phosphatase.

Article Snippet: Immunohistochemical staining was performed using primary antibodies against osteocalcin (OCN) (Cat no.: GB11233, Servicebio, China) with a dilution of 1:300, tumor necrosis factor-alpha (TNF-α) (Cat no.: GB11188, Servicebio, China) with a dilution of 1:800, and interleukin-1 beta (IL-1β) (Cat no.: GB11113, Servicebio, China) with a dilution of 1:400.

Techniques:

Journal: Biomaterials Translational

Article Title: Ångstrom-scale silver particle-infused hydrogels eliminate orthopedic implant infections and support fracture healing

doi: 10.12336/biomatertransl.2025.01.007

Figure Lengend Snippet: Note: Statistical significance determined at * p < 0.05 and ** p < 0.01. Abbreviations: AgÅPs: Ångstrom-scale silver particles; E. coli: Escherichia coli ; Gel: Hydrogel; IM: Induced medium ; OCN: Osteocalcin; TRAP: Tartrate resistant acid phosphatase.

Article Snippet: Immunohistochemical staining was performed using primary antibodies against osteocalcin (OCN) (Cat no.: GB11233, Servicebio, China) with a dilution of 1:300, tumor necrosis factor-alpha (TNF-α) (Cat no.: GB11188, Servicebio, China) with a dilution of 1:800, and interleukin-1 beta (IL-1β) (Cat no.: GB11113, Servicebio, China) with a dilution of 1:400.

Techniques: