primary antibodies targeting col-ii and ocn  (Proteintech)

Journal: Materials Today Bio

Article Title: Icariin-loaded GelMa hydrogel encapsulated potassium sodium niobate biomimetic piezoelectric scaffold regulates macrophage polarization to accelerate bone defect repair

doi: 10.1016/j.mtbio.2025.102476

Figure Lengend Snippet: Ultrasound-dependent osteogenic differentiation of BMSCs on composite scaffolds. (A)Alkaline phosphatase (ALP) staining after 7 days showing early osteogenic differentiation. Purple staining indicates ALP activity. (B) Alizarin Red S (ARS) staining after 14 days showing mineral deposition. Orange-red staining indicates calcium phosphate precipitation. (C) Quantitative analysis of ALP activity normalized to OD450. (D) Quantitative analysis of mineral deposition by ARS absorption at 570 nm. (E) Western blot analysis of COL1 and RUNX2 proteins with β-actin loading control. (F, G) Quantitative protein expression analysis of COL1 (F) and RUNX2 (G) normalized to β-actin. (H) Heat map showing fold-change in osteogenic gene expression (ALP, RUNX2, BMP2, COL1, OPN, OCN) under different conditions. Data represent mean ± SD (n = 6). Statistical significance: ∗∗p < 0.01, ∗∗∗p < 0.001 comparing US(+) vs US(−) within the same group. US(−): static culture; US(+): ultrasonic stimulation (1.5 MHz, 30 mW/cm 2 , 10 min/day).

Article Snippet: At 12 weeks post-surgery, immunohistochemistry was used to assess new bone formation using anti-bone sialoprotein (BSP) and anti-osteocalcin (OCN) primary antibodies (Servicebio, Wuhan, China; 1:200 dilution) incubated overnight at 4 °C, and rabbit IgG secondary antibodies (2-step plus Poly-HRP Anti Rabbit IgG Detection System) incubated at room temperature for 2 h.

Techniques: Staining, Activity Assay, Western Blot, Control, Expressing, Gene Expression

Journal: Materials Today Bio

Article Title: Icariin-loaded GelMa hydrogel encapsulated potassium sodium niobate biomimetic piezoelectric scaffold regulates macrophage polarization to accelerate bone defect repair

doi: 10.1016/j.mtbio.2025.102476

Figure Lengend Snippet: ICA@G/NHP scaffolds promote M2 macrophage polarization through C-type lectin receptor signaling pathway. (A) Flow cytometric analysis of CD86 (M1 marker) and CD206 (M2 marker) expression in RAW264.7 macrophages after 48h treatment with different scaffold formulations. (B) Quantitative analysis of CD206-positive cell percentages showing significant M2 polarization in ICA@G and ICA@G/NHP groups. (C–D) Western blot analysis and quantification of M2 markers (CD163, Arg-1) and M1 marker (iNOS) confirming phenotypic shifts toward M2 polarization. (E) Schematic illustration of macrophage-conditioned medium treatment protocol for BMSC osteogenic differentiation assessment. (F–I) qPCR analysis of osteogenic markers (COL1, OCN, RUNX2, ALP) in BMSCs cultured with macrophage-conditioned medium, demonstrating enhanced osteogenic differentiation. (J–K) Network pharmacology analysis showing enrichment of C-type lectin receptor signaling pathway and transcriptional regulation processes. (L–M) Western blot analysis and quantification of C-type lectin receptor pathway components (Dectin-1, Syk, phospho-Syk, STAT6) revealing mechanistic basis for icariin-mediated M2 polarization. Data presented as mean ± SD (n = 3), ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: At 12 weeks post-surgery, immunohistochemistry was used to assess new bone formation using anti-bone sialoprotein (BSP) and anti-osteocalcin (OCN) primary antibodies (Servicebio, Wuhan, China; 1:200 dilution) incubated overnight at 4 °C, and rabbit IgG secondary antibodies (2-step plus Poly-HRP Anti Rabbit IgG Detection System) incubated at room temperature for 2 h.

Techniques: Marker, Expressing, Western Blot, Cell Culture

Journal: Materials Today Bio

Article Title: Icariin-loaded GelMa hydrogel encapsulated potassium sodium niobate biomimetic piezoelectric scaffold regulates macrophage polarization to accelerate bone defect repair

doi: 10.1016/j.mtbio.2025.102476

Figure Lengend Snippet: (A) IHC for OCN and BSP at the 6- and 12-week time point, respectively. (B, C) Quantitative analysis of OCN and BSP expression positive areas by using Image J software.

Article Snippet: At 12 weeks post-surgery, immunohistochemistry was used to assess new bone formation using anti-bone sialoprotein (BSP) and anti-osteocalcin (OCN) primary antibodies (Servicebio, Wuhan, China; 1:200 dilution) incubated overnight at 4 °C, and rabbit IgG secondary antibodies (2-step plus Poly-HRP Anti Rabbit IgG Detection System) incubated at room temperature for 2 h.

Techniques: Expressing, Software

Journal: Gut Microbes

Article Title: Intermittent fasting triggers interorgan communication to improve the progression of diabetic osteoporosis

doi: 10.1080/19490976.2025.2555619

Figure Lengend Snippet: Intermittent fasting intervention modulates gut microbiota alterations in diabetic mice. (A) Representative diagram of HE stain, trap stain, and OCN immunohistochemistry stain.(B)N.Obs/BS.(C)ostrocalcin.(D)MAR.(E)BFR.(F)maximum force. (G) stiffness. (H)community barplot analysis. (I)principal cause analysis.(J)Typing analysis on phylum level. (K)microbiotal dysbiosis index. (L) correlation analysis of intestinal microbiota.(M) heatmap analysis.(N) feature enrichment analysis.(O) venn plot. (P) COG function classification. Data are presented as mean ± SEM and statistical significance was determined by two-way ANOVA with Newman-Keuls multiple comparisons test, n = 12 mice per group, * p < 0.05, ** p < 0.01.

Article Snippet: The osteocalcin (OCN) primary antibody (Cat. No. gb11233; 1:200) and secondary antibody (Cat. No. gb23303; 1:200) were provided by Servicebio (Wuhan, China).

Techniques: H&E Stain, Staining, Immunohistochemistry

Journal: Gut Microbes

Article Title: Intermittent fasting triggers interorgan communication to improve the progression of diabetic osteoporosis

doi: 10.1080/19490976.2025.2555619

Figure Lengend Snippet: Effects of Chr supplementation on diabetes-induced osteoporosis. (A) Representative diagram of bone microstructure, HE stain, OCN stain. (B)BMD of the fourth lumbar vertebrae. (C)BV/TV/BW of the fourth lumbar vertebrae. (D)tb.Th of fourth lumbar vertebrae. (E)tb.N of fourth lumbar vertebrae. (F)tb.Sp of the fourth lumbar vertebrae. (G) Tb.Th of femur. (H)tb.N of femur. (I)tb.Sp of femur. (J) BV/TV/BW of femur. (K)length of femur. (T) Ct.Th femur. (U) Ct.Ar/Tt.Ar of femur. (O)CD4+ T lymphocytes. (P) histogram of intestinal microbiota composition. (Q)the total load of gut microbiota. (R) Escherichia coli relative abundance. (S)christensenellaceae relative abundance. Data are presented as mean ± SEM, and statistical significance was determined by two-way ANOVA with Newman-Keuls multiple comparisons test, n = 12 mice per group, * p < 0.05, ** p < 0.01.

Article Snippet: The osteocalcin (OCN) primary antibody (Cat. No. gb11233; 1:200) and secondary antibody (Cat. No. gb23303; 1:200) were provided by Servicebio (Wuhan, China).

Techniques: H&E Stain, Staining

Journal: Gut Microbes

Article Title: Intermittent fasting triggers interorgan communication to improve the progression of diabetic osteoporosis

doi: 10.1080/19490976.2025.2555619

Figure Lengend Snippet: Effects of fecal transplantation assays and gut microbiota elimination assays on diabetes-mediated osteoporosis. (A) Representative diagram of bone microstructure, HE stain, OCN stain of fecal transplantation assays, and gut microbiota elimination experiment. (B)tb.Sp of the fourth lumbar vertebrae. (C)tb.Th of fourth lumbar vertebrae. (D)tb.N of fourth lumbar vertebrae. (E)BV/TV/BW of the fourth lumbar vertebrae. (F)BMD of the fourth lumbar vertebrae. (G)tb.Sp of femur.(H) Tb.Th femur. (K)TRAP expression. (L)OCN expression. (M) femur BMD in gut microbiota elimination assays. (N) BV/TV/BW of femur in gut microbiota elimination assays. (O)tb.N of femur in gut microbiota elimination assays. (P) Tb.undefined.The femur of in gut microbiota elimination assays. (Q) Tb.Sp of femur in gut microbiota elimination assays. (R) BMD of the fourth lumbar vertebrae in gut microbiota elimination assays. (S)serum LPS. (T) correlated BV/TV with LPS level. Data are presented as mean ± SEM, and statistical significance was determined by two-way ANOVA with Newman-Keuls multiple comparisons test, n = 12 mice per group, * p < 0.05, ** p < 0.01.

Article Snippet: The osteocalcin (OCN) primary antibody (Cat. No. gb11233; 1:200) and secondary antibody (Cat. No. gb23303; 1:200) were provided by Servicebio (Wuhan, China).

Techniques: Transplantation Assay, H&E Stain, Staining, Expressing

Journal: Gut Microbes

Article Title: Intermittent fasting triggers interorgan communication to improve the progression of diabetic osteoporosis

doi: 10.1080/19490976.2025.2555619

Figure Lengend Snippet: Chr source mir-551b in the regulation of osteoclast activity. (A) confocal microscopy analysis of the femoral sections from mice treated with the PKH67-labeled EVs for 1 h by oral route(top line) and Representative images of the Chr -EVs antibody (ab)-stained(bottom line). (B-C) quantification of the fluorescent signals in PKH67-labeled EVs and Chr -EVs antibody (ab)-stained. (E) Principal cause analysis of miRNA analysis. (F) volcano map miRNA analysis. (G) KEGG. (H) heatmap of miRNA analysis. (I) relative of four miRNA levels. (J) relative of miR-551b level in EV. (K)relative of miRNA-551b level after depletion of EV. (L) relative of miR-551b level after treatment with miR-551b mimics and inhibitors.(M)ALP activity. (N) OD value. (O) relative OCN mRNA. (P)relative ALP mRNA. Data are presented as mean ± SEM, and statistical significance was determined by two-way ANOVA with Newman-Keuls multiple comparisons test, n = 12 mice per group, * p < 0.05, ** p < 0.01.

Article Snippet: The osteocalcin (OCN) primary antibody (Cat. No. gb11233; 1:200) and secondary antibody (Cat. No. gb23303; 1:200) were provided by Servicebio (Wuhan, China).

Techniques: Activity Assay, Confocal Microscopy, Labeling, Staining

Journal: Bioactive Materials

Article Title: Pretreated exosomes by electrical stimulation accelerate bone regeneration

doi: 10.1016/j.bioactmat.2025.04.019

Figure Lengend Snippet: Histological staining of the distal femur defect site at 6-weeks post-injection. (A) Hematoxylin and eosin (H&E) staining, (B) Masson's trichrome staining, and (C) Immunohistochemical staining for OCN. D) The relative proportion of collagen from the quantitative analysis of Masson staining. E) Quantitative analysis results of IHC Results are shown as mean ± SD. ns: non-significant differences, ∗∗p < 0.01.

Article Snippet: They were then co-incubated with the primary antibody, specifically the OCN primary antibody from Beyotime, China.

Techniques: Staining, Injection, Immunohistochemical staining