elisa mouse ocn eia kit  (Biomedical Technologies)

Journal: Experimental & Molecular Medicine

Article Title: Hypoxia-inducible factor-2α mediates senescence-associated intrinsic mechanisms of age-related bone loss

doi: 10.1038/s12276-021-00594-y

Figure Lengend Snippet: a Detection of the indicated mRNAs in the long bones of 1.5-, 4-, and 20-month-old mice. The levels of mRNAs encoded by Hif-2α , osteoblast marker genes ( Ocn and Runx2 ), an osteoclast marker gene ( Trap ) and senescence marker genes ( p16 and p21 ) were determined by RT-PCR and quantified by qRT-PCR ( n ≥ 4). b The protein levels of HIF-2α extracted from mice of the indicated age determined by western blot analysis (left panel) and quantified by a CS analyzer 4 (right panel) ( n = 5). c ELISA-based measurements of the serum concentrations of OCN ( n = 5) and CTX-1 ( n = 5) in 1.5-, 4-, 12- and 20-month-old mice. d, e Immunostaining of osteoblasts ( d ) and osteoclasts ( e ) from the bone tissues of young (4-month-old) and old (12-month-old) mice for HIF-2α. The dotted lines indicate osteoblasts, and the arrowheads indicate osteoclasts. Scale bar: 25 μm. Quantification of HIF-2α-positive cells is shown in the right panel ( n = 8). M = month. The values are presented as the means ± SDs (* P < 0.05, ** P < 0.01, and *** P < 0.005).

Article Snippet: Serum OCN and CTX-1 levels were measured using a mouse Gla-OCN High Sensitive EIA Kit (MK127; TaKaRa Bio) and CTX-1 EIA Kit (NBP2-69074; Novus Biologicals).

Techniques: Marker, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Immunostaining

Journal: Experimental & Molecular Medicine

Article Title: Hypoxia-inducible factor-2α mediates senescence-associated intrinsic mechanisms of age-related bone loss

doi: 10.1038/s12276-021-00594-y

Figure Lengend Snippet: a , b Doxorubicin-induced inhibition of osteoblast differentiation. Primary calvarial preosteoblasts from newborn mice were cultured in osteogenic differentiation medium (DM) containing 50 μg/ml L-AA and 5 mM β-GP for 7 days. Before harvesting cells on day 14, 0.01 μg/ml doxorubicin was added to the differentiation medium for 1, 3, or 5 days. Representative images of alkaline phosphatase (ALP) and alizarin red S (ARS) staining in primary calvarial preosteoblasts cultured in control medium (CM) or differentiation medium in the absence or presence of 0.01 μg/ml doxorubicin are shown ( a ). The expression levels of Hif-2α, Ocn, Runx2, p16 , and p21 were analyzed by qRT-PCR ( n = 5; b ). c Quantification of mRNA levels by qRT-PCR. The transcript levels of the indicated genes were measured in primary calvarial preosteoblasts infected with Ad-C at an MOI of 800 or Ad -Hif-2α at the indicated MOI ( n ≥ 4). d The mRNA levels of Hif-2α, Ocn, Runx2, p16 , and p21 . Osteoblasts isolated from Hif-2α fl/fl mice were infected with Ad-C or Ad- Cre in the presence of differentiation medium ( n ≥ 4). e , f Effect of HIF-2α inhibition on osteoblast matrix maturation. Doxorubicin-treated osteoblasts were treated with the potent HIF-2α inhibitor TC-S 7009 and incubated for 3 days. The levels of Ocn , Runx2, p16 , and p21 mRNAs were measured by qRT-PCR ( n = 5; e ). Assessment of osteoblast differentiation and mineral deposition by ALP and ARS staining ( f ). The values are presented as the means ± SDs (* P < 0.05, ** P < 0.01, and *** P < 0.005; NS not significant).

Article Snippet: Serum OCN and CTX-1 levels were measured using a mouse Gla-OCN High Sensitive EIA Kit (MK127; TaKaRa Bio) and CTX-1 EIA Kit (NBP2-69074; Novus Biologicals).

Techniques: Inhibition, Cell Culture, Staining, Expressing, Quantitative RT-PCR, Infection, Isolation, Incubation

Journal: Frontiers in Pharmacology

Article Title: Osteogenic Enhancement Between Icariin and Bone Morphogenetic Protein 2: A Potential Osteogenic Compound for Bone Tissue Engineering

doi: 10.3389/fphar.2019.00201

Figure Lengend Snippet: Primer sequences for real-time quantitative polymerase chain reaction analysis of the gene expression.

Article Snippet: The supernatant OCN concentration was determined using the ELISA mouse OCN EIA kit (Biomedical Technologies, Stoughton, MA, United States).

Techniques: Real-time Polymerase Chain Reaction, Expressing

Journal: Frontiers in Pharmacology

Article Title: Osteogenic Enhancement Between Icariin and Bone Morphogenetic Protein 2: A Potential Osteogenic Compound for Bone Tissue Engineering

doi: 10.3389/fphar.2019.00201

Figure Lengend Snippet: Osteogenic differentiation results of MC3T3-E1 cells after treatment with BioCaP alone or BioCaP with Icariin or/and BMP-2. (A) The ALP activities were determined by colorimetric assay on days 1, 4, and 7. (B) The OCN expressions on days 4, 7, and 10. (C) Macroscopic images of alzarin red stained 3, 4, and 5 weeks after five groups treatment. (D) The mineralization results on 3, 4, and 5 weeks. Mean values ( n = 3 samples per group) were represented together with the standard deviation normalized by total cellular protein. Error bars denote the standard deviation. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗ without lines means versus control (C). C, Control; BioCaP, biomimetic calcium phosphate; I, Icariin; B, BMP-2.

Article Snippet: The supernatant OCN concentration was determined using the ELISA mouse OCN EIA kit (Biomedical Technologies, Stoughton, MA, United States).

Techniques: Colorimetric Assay, Staining, Standard Deviation, Control

Journal: Frontiers in Pharmacology

Article Title: Osteogenic Enhancement Between Icariin and Bone Morphogenetic Protein 2: A Potential Osteogenic Compound for Bone Tissue Engineering

doi: 10.3389/fphar.2019.00201

Figure Lengend Snippet: The time course changes in mRNA expression of (A) ALP, (B) BMP-2 (C) Col1, (D) OCN, (E) Runx2 in MC3T3-E1 on days 1, 4, and 7. Mean values ( n = 3 samples per group) were represented together with the standard deviation. Error bars denote the standard deviation. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗ without lines means versus control (C). C, Control; BioCaP, biomimetic calcium phosphate; I, Icariin; B, BMP-2.

Article Snippet: The supernatant OCN concentration was determined using the ELISA mouse OCN EIA kit (Biomedical Technologies, Stoughton, MA, United States).

Techniques: Expressing, Standard Deviation, Control

Journal: Frontiers in Pharmacology

Article Title: Osteogenic Enhancement Between Icariin and Bone Morphogenetic Protein 2: A Potential Osteogenic Compound for Bone Tissue Engineering

doi: 10.3389/fphar.2019.00201

Figure Lengend Snippet: Factor in mRNA expression as a function of the treatment and of the days after treatment.

Article Snippet: The supernatant OCN concentration was determined using the ELISA mouse OCN EIA kit (Biomedical Technologies, Stoughton, MA, United States).

Techniques: Expressing, Control