primary antibody for osteocalcin  (Proteintech)

Journal: Bioactive Materials

Article Title: Dual-function thermoresponsive antibiotic-loaded hydrogel with antimicrobial and osteogenic properties for implant-related infection control

doi: 10.1016/j.bioactmat.2026.02.044

Figure Lengend Snippet: In vivo immunohistochemical analysis (n = 6). (A) Representative immunolabeling images of the Control, PNVCL, and PNVCL/TC 25 mg/mL groups showing osteopontin (OPN) and osteocalcin (OCN) expression (immunopositive cells indicated by black arrows). (B) Mean scores (0–3) ± standard deviation for OPN immunostaining. (C) Mean scores (0–3) ± standard deviation for OCN immunostaining. Bars indicate statistically significant differences between groups (p < 0.05; Tukey HSD test).

Article Snippet: Endogenous peroxidase activity was quenched by incubation with 3% hydrogen peroxide for 1 h, followed by blocking of nonspecific binding sites with 1% bovine serum albumin for 12 h. The sections were then incubated with goat anti-osteopontin and goat anti-osteocalcin primary antibodies (sc-21742 and sc-30044, respectively; Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: In Vivo, Immunohistochemical staining, Immunolabeling, Control, Expressing, Standard Deviation, Immunostaining

Journal: Bioactive Materials

Article Title: Ribbon-shaped microgels as bioinks for 3D bioprinting of anisotropic tissue structures

doi: 10.1016/j.bioactmat.2025.12.040

Figure Lengend Snippet: μRB bioinks modulate MSC morphology, osteogenesis and bone formation in a stiffness-dependent manner. (A) Live/dead staining of MSCs after extrusion (Day 0) or after 28 days of culture in osteogenic medium. Green: live cells, Red: dead cells. (B) Metabolic activity of MSCs measured by PrestoBlue assay at day 0 and day 14 after bioprinting (n = 6 per group). (C) DNA content per scaffold measured by PicoGreen assay at day 28 (n = 3 per group). (D) Alizarin red S (ARS) staining for mineralized bone matrix, (E) Aniline Blue staining for total collagen, and (F) immunostaining of Osteocalcin (OCN), a mature bone marker, at day 14 and day 21 of osteogenesis (n = 4 per group). (G–I) Quantification of ARS and Aniline Blue percent positive area and OCN mean fluorescence intensity (MFI). Scale bar = 100 μm. Values are presented as mean ± S.D. and p-values were determined by one-way analysis of variance (ANOVA) with Tukey's multiple comparisons test; ∗p ≤ 0.05, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.005, ∗∗∗∗p ≤ 0.001.

Article Snippet: Primary antibody for osteocalcin (1:200, 23418-1-AP, Proteintech), GFP (1:100, 50430-2-AP, Proteintech) was diluted in 1 % BSA with 0.1 % Triton X-100 and incubated overnight at 4 °C.

Techniques: Staining, Activity Assay, Prestoblue Assay, Picogreen Assay, Immunostaining, Marker, Fluorescence

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