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alp staining buffer  (Beyotime)


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    Beyotime alp staining buffer
    ( A ) Micro-CT scan and 3D reconstruction results (scale bar, 1 mm); quantitative analysis of bone density, volume, and other parameters. BV/TV, bone tissue volume/total tissue; Tb.Th, trabecular thickness; Tb.N, trabecular number. ( B ) H&E staining images of different groups (scale bar, 1 mm and 125 μm). ( C ) Masson staining images of different groups (scale bar, 1 mm and 125 μm). ( D ) Flow cytometry <t>showed</t> <t>BMSCs</t> from osteoporotic rats were positive for CD44 (99.8%), CD29 (99.8%), CD73 (98.9%), and CD90 (77.3%) and negative for CD34 (2.57%) and CD45 (2.5%). ( E ) Alizarin Red S, Toluidine blue, and Oil Red O staining were used to detect osteogenic, chondrogenic, and adipogenic differentiation, respectively (scale bar, 100 μm). ( F ) SA-β-gal staining detected senescent cells (scale bar, 100 μm). ( G ) Western blotting analysis of Nanog, Sox2, and Oct4 protein expression in BMSCs from different groups. ( H ) Western blotting analysis of P53, P21, and P16 protein expression in BMSCs from different groups. ( I ) Representative immunofluorescence staining of γ-H2AX foci formation in BMSCs from different groups (scale bar, 25 μm). DAPI, 4′,6-diamidino-2-phenylindole. ( J ) Representative immunofluorescence staining of CD90 and Runx2 in BMSCs from the different groups (scale bar, 25 μm). ( K ) Representative immunofluorescence staining of CD90 and Opn in BMSCs from different groups (scale bar, 25 μm). ( L ) Western blotting analysis of Runx2 and Opn protein expression in BMSCs from different groups. ( M ) <t>ALP</t> staining of BMSCs from the different groups. ( N ) Alizarin Red S staining of BMSCs from different groups. n = 3 for each group. Error bars denote means ± SEM; ns, no significance; * P < 0.05, ** P < 0.01, and *** P < 0.001.
    Alp Staining Buffer, supplied by Beyotime, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/alp+staining+buffer/alp+staining+buffer/pmc11529719-242-12-15
    Average 90 stars, based on 1 article reviews
    alp staining buffer - by Bioz Stars, 2026-09
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    Images

    1) Product Images from "Stem cell–homing biomimetic hydrogel promotes the repair of osteoporotic bone defects through osteogenic and angiogenic coupling"

    Article Title: Stem cell–homing biomimetic hydrogel promotes the repair of osteoporotic bone defects through osteogenic and angiogenic coupling

    Journal: Science Advances

    doi: 10.1126/sciadv.adq6700

    ( A ) Micro-CT scan and 3D reconstruction results (scale bar, 1 mm); quantitative analysis of bone density, volume, and other parameters. BV/TV, bone tissue volume/total tissue; Tb.Th, trabecular thickness; Tb.N, trabecular number. ( B ) H&E staining images of different groups (scale bar, 1 mm and 125 μm). ( C ) Masson staining images of different groups (scale bar, 1 mm and 125 μm). ( D ) Flow cytometry showed BMSCs from osteoporotic rats were positive for CD44 (99.8%), CD29 (99.8%), CD73 (98.9%), and CD90 (77.3%) and negative for CD34 (2.57%) and CD45 (2.5%). ( E ) Alizarin Red S, Toluidine blue, and Oil Red O staining were used to detect osteogenic, chondrogenic, and adipogenic differentiation, respectively (scale bar, 100 μm). ( F ) SA-β-gal staining detected senescent cells (scale bar, 100 μm). ( G ) Western blotting analysis of Nanog, Sox2, and Oct4 protein expression in BMSCs from different groups. ( H ) Western blotting analysis of P53, P21, and P16 protein expression in BMSCs from different groups. ( I ) Representative immunofluorescence staining of γ-H2AX foci formation in BMSCs from different groups (scale bar, 25 μm). DAPI, 4′,6-diamidino-2-phenylindole. ( J ) Representative immunofluorescence staining of CD90 and Runx2 in BMSCs from the different groups (scale bar, 25 μm). ( K ) Representative immunofluorescence staining of CD90 and Opn in BMSCs from different groups (scale bar, 25 μm). ( L ) Western blotting analysis of Runx2 and Opn protein expression in BMSCs from different groups. ( M ) ALP staining of BMSCs from the different groups. ( N ) Alizarin Red S staining of BMSCs from different groups. n = 3 for each group. Error bars denote means ± SEM; ns, no significance; * P < 0.05, ** P < 0.01, and *** P < 0.001.
    Figure Legend Snippet: ( A ) Micro-CT scan and 3D reconstruction results (scale bar, 1 mm); quantitative analysis of bone density, volume, and other parameters. BV/TV, bone tissue volume/total tissue; Tb.Th, trabecular thickness; Tb.N, trabecular number. ( B ) H&E staining images of different groups (scale bar, 1 mm and 125 μm). ( C ) Masson staining images of different groups (scale bar, 1 mm and 125 μm). ( D ) Flow cytometry showed BMSCs from osteoporotic rats were positive for CD44 (99.8%), CD29 (99.8%), CD73 (98.9%), and CD90 (77.3%) and negative for CD34 (2.57%) and CD45 (2.5%). ( E ) Alizarin Red S, Toluidine blue, and Oil Red O staining were used to detect osteogenic, chondrogenic, and adipogenic differentiation, respectively (scale bar, 100 μm). ( F ) SA-β-gal staining detected senescent cells (scale bar, 100 μm). ( G ) Western blotting analysis of Nanog, Sox2, and Oct4 protein expression in BMSCs from different groups. ( H ) Western blotting analysis of P53, P21, and P16 protein expression in BMSCs from different groups. ( I ) Representative immunofluorescence staining of γ-H2AX foci formation in BMSCs from different groups (scale bar, 25 μm). DAPI, 4′,6-diamidino-2-phenylindole. ( J ) Representative immunofluorescence staining of CD90 and Runx2 in BMSCs from the different groups (scale bar, 25 μm). ( K ) Representative immunofluorescence staining of CD90 and Opn in BMSCs from different groups (scale bar, 25 μm). ( L ) Western blotting analysis of Runx2 and Opn protein expression in BMSCs from different groups. ( M ) ALP staining of BMSCs from the different groups. ( N ) Alizarin Red S staining of BMSCs from different groups. n = 3 for each group. Error bars denote means ± SEM; ns, no significance; * P < 0.05, ** P < 0.01, and *** P < 0.001.

    Techniques Used: Micro-CT, Staining, Flow Cytometry, Western Blot, Expressing, Immunofluorescence

    ( A ) Immunofluorescence staining of F-actin in BMSCs cultured with hydrogels for 7 days (scale bar, 25 μm). ( B ) Live/dead assay of BMSCs cultured with hydrogels for 3 days (scale bar, 100 μm). Live cells appear green, and dead cells appear red. ( C ) Western blots analysis of P-eNOS and eNOS levels in BMSCs cultured with hydrogels for 7 days. ( D ) NO generation in BMSCs cultured with hydrogels for 7 days indicated by nitrite levels. ( E ) cGMP expression in BMSCs cultured with hydrogels for 7 days. ( F ) Western blots analysis of sGC and PKG levels in BMSCs cultured with hydrogels for 7 days. ( G ) Western blots analysis of Runx2 and Opn levels in BMSCs cultured with hydrogels for 7 days. ( H ) Representative of immunofluorescence staining of CD90 and Runx2 in BMSCs cultured with hydrogels for 7 days (scale bar, 25 μm). ( I ) Representative of immunofluorescence staining of CD90 and Opn in BMSCs cultured with hydrogels for 7 days (scale bar, 25 μm). ( J ) The ALP staining of BMSCs cultured with hydrogels for 7 days. ( K ) Alizarin Red S staining of BMSCs cultured with hydrogels for 7 days. ( L ) Schematic diagram of SA-MSNs@CM-Stiff activating NO/cGMP pathway to promote BMSC osteogenesis. n = 3 for each group. Error bars denote means ± SEM; * P < 0.05, ** P < 0.01, and *** P < 0.001.
    Figure Legend Snippet: ( A ) Immunofluorescence staining of F-actin in BMSCs cultured with hydrogels for 7 days (scale bar, 25 μm). ( B ) Live/dead assay of BMSCs cultured with hydrogels for 3 days (scale bar, 100 μm). Live cells appear green, and dead cells appear red. ( C ) Western blots analysis of P-eNOS and eNOS levels in BMSCs cultured with hydrogels for 7 days. ( D ) NO generation in BMSCs cultured with hydrogels for 7 days indicated by nitrite levels. ( E ) cGMP expression in BMSCs cultured with hydrogels for 7 days. ( F ) Western blots analysis of sGC and PKG levels in BMSCs cultured with hydrogels for 7 days. ( G ) Western blots analysis of Runx2 and Opn levels in BMSCs cultured with hydrogels for 7 days. ( H ) Representative of immunofluorescence staining of CD90 and Runx2 in BMSCs cultured with hydrogels for 7 days (scale bar, 25 μm). ( I ) Representative of immunofluorescence staining of CD90 and Opn in BMSCs cultured with hydrogels for 7 days (scale bar, 25 μm). ( J ) The ALP staining of BMSCs cultured with hydrogels for 7 days. ( K ) Alizarin Red S staining of BMSCs cultured with hydrogels for 7 days. ( L ) Schematic diagram of SA-MSNs@CM-Stiff activating NO/cGMP pathway to promote BMSC osteogenesis. n = 3 for each group. Error bars denote means ± SEM; * P < 0.05, ** P < 0.01, and *** P < 0.001.

    Techniques Used: Immunofluorescence, Staining, Cell Culture, Live Dead Assay, Western Blot, Expressing

    Related Articles

    Incubation:

    Article Title: Stem cell–homing biomimetic hydrogel promotes the repair of osteoporotic bone defects through osteogenic and angiogenic coupling
    Article Snippet: .. Following two washes in PBS, BMSCs were incubated for 6 hours in ALP staining buffer (Beyotime Institute of Biotechnology, China). ..

    Article Title: The role of ZBTB16 in odontogenic differentiation of dental pulp stem cells.
    Article Snippet: Following this, the cells were washed with PBS twice and fixed using 4% Paraformaldehyde Fix Solution (PFA; Sangon Biotech, Shanghai, China) for 30 min. .. Cells were incubated with ALP staining buffer (Beyotime, Shanghai, China), which was prepared according to the manufacturer’s instructions, for 20–60 min at room temperature, after which the cells were washed with PBS to stop the reaction. .. The ALP activity was measured using Alkaline Phosphatase Assay Kit (Beyotime) according to the manufacturer’s instructions after cells were lysed with buffer containing 0.1% Triton X100.

    Article Title: Targeting EZH2 attenuates the ferroptosis-mediated osteoblast-osteoclast imbalance in rheumatoid arthritis.
    Article Snippet: Objective: The enhancer of zeste 2 polycomb repressive complex 2 subunit (EZH2) can regulate osteogenesis and osteoclastogenesis.. This study aimed to further explore the effects of EZH2 modification on ferroptosis and the osteoblast–osteoclast balance in rheumatoid arthritis (RA) in vitro and in vivo.. Methods: Bone marrow mesenchymal stromal cells were transfected with EZH2 overexpression (oeEZH2) and EZH2 shRNA (shEZH2) plasmids with or without ferrostatin-1 (Fer-1) treatment and subjected to an osteoblast differentiation assay.

    Article Title: Damaged brain accelerates bone healing by releasing small extracellular vesicles that target osteoprogenitors
    Article Snippet: .. BMSC and MC3T3-E1 cells were washed with PBS twice and then incubated with ALP staining buffer (Beyotime Institute of Biotechnology, Beijing, China) at 37 °C in the dark for 30 min. Then, the reaction was stopped by using distilled water, and the plate was dried before taking photographic images. ..

    Staining:

    Article Title: Stem cell–homing biomimetic hydrogel promotes the repair of osteoporotic bone defects through osteogenic and angiogenic coupling
    Article Snippet: .. Following two washes in PBS, BMSCs were incubated for 6 hours in ALP staining buffer (Beyotime Institute of Biotechnology, China). ..

    Article Title: Enhanced effects of antagomiR-3074-3p-conjugated PEI-AuNPs on the odontogenic differentiation by targeting FKBP9.
    Article Snippet: .. After 7 days of mineralization induction, hDPSCs were washed with 1× PBS and fixed with 4% paraformaldehyde for 15 min. Then, cells were washed several times with deionized water and treated with prepared ALP staining buffer (Beyotime, Shanghai, China) at 37°C for 30 min. After washing several times with 1× PBS, the intensity of dark blue was assessed under an inverted microscope (Leica, Germany). ..

    Article Title: Precisely Tuning the Pore-Wall Surface Composition of Bioceramic Scaffolds Facilitates Angiogenesis and Orbital Bone Defect Repair.
    Article Snippet: Orbital bone damage (OBD) may result in severe post-traumatic enophthalmos, craniomaxillofacial deformities, vision loss, and intracranial infections.. However, it is still a challenge to fabricate advanced biomaterials that can match the individual anatomical structure and enhance OBD repair in situ.. Herein, we aimed to develop a selective surface modification strategy on bioceramic scaffolds and evaluated the effects of inorganic or organic functional coating on angiogenesis and osteogenesis, ectopically and orthotopically in OBD models.

    Article Title: Baicalein Accelerates Tendon-Bone Healing via Activation of Wnt/ β -Catenin Signaling Pathway in Rats.
    Article Snippet: ALP activity was checked using a fluorometric detection kit according to the instructions (Nanjing Jiancheng Biotechnology Co., Ltd., Jiangsu, China). .. According to the instructions of ALP staining (Beyotime Biotechnology), parallel wells were added to ALP staining buffer to observe the quantity of the ALP. .. The results were observed by taking photos with a microscope (Olympus Corporation, Tokyo, Japan).

    Article Title: The role of ZBTB16 in odontogenic differentiation of dental pulp stem cells.
    Article Snippet: Following this, the cells were washed with PBS twice and fixed using 4% Paraformaldehyde Fix Solution (PFA; Sangon Biotech, Shanghai, China) for 30 min. .. Cells were incubated with ALP staining buffer (Beyotime, Shanghai, China), which was prepared according to the manufacturer’s instructions, for 20–60 min at room temperature, after which the cells were washed with PBS to stop the reaction. .. The ALP activity was measured using Alkaline Phosphatase Assay Kit (Beyotime) according to the manufacturer’s instructions after cells were lysed with buffer containing 0.1% Triton X100.

    Article Title: Targeting EZH2 attenuates the ferroptosis-mediated osteoblast-osteoclast imbalance in rheumatoid arthritis.
    Article Snippet: Objective: The enhancer of zeste 2 polycomb repressive complex 2 subunit (EZH2) can regulate osteogenesis and osteoclastogenesis.. This study aimed to further explore the effects of EZH2 modification on ferroptosis and the osteoblast–osteoclast balance in rheumatoid arthritis (RA) in vitro and in vivo.. Methods: Bone marrow mesenchymal stromal cells were transfected with EZH2 overexpression (oeEZH2) and EZH2 shRNA (shEZH2) plasmids with or without ferrostatin-1 (Fer-1) treatment and subjected to an osteoblast differentiation assay.

    Article Title: Damaged brain accelerates bone healing by releasing small extracellular vesicles that target osteoprogenitors
    Article Snippet: .. BMSC and MC3T3-E1 cells were washed with PBS twice and then incubated with ALP staining buffer (Beyotime Institute of Biotechnology, Beijing, China) at 37 °C in the dark for 30 min. Then, the reaction was stopped by using distilled water, and the plate was dried before taking photographic images. ..

    Inverted Microscopy:

    Article Title: Enhanced effects of antagomiR-3074-3p-conjugated PEI-AuNPs on the odontogenic differentiation by targeting FKBP9.
    Article Snippet: .. After 7 days of mineralization induction, hDPSCs were washed with 1× PBS and fixed with 4% paraformaldehyde for 15 min. Then, cells were washed several times with deionized water and treated with prepared ALP staining buffer (Beyotime, Shanghai, China) at 37°C for 30 min. After washing several times with 1× PBS, the intensity of dark blue was assessed under an inverted microscope (Leica, Germany). ..



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    Image Search Results


    ( A ) Micro-CT scan and 3D reconstruction results (scale bar, 1 mm); quantitative analysis of bone density, volume, and other parameters. BV/TV, bone tissue volume/total tissue; Tb.Th, trabecular thickness; Tb.N, trabecular number. ( B ) H&E staining images of different groups (scale bar, 1 mm and 125 μm). ( C ) Masson staining images of different groups (scale bar, 1 mm and 125 μm). ( D ) Flow cytometry showed BMSCs from osteoporotic rats were positive for CD44 (99.8%), CD29 (99.8%), CD73 (98.9%), and CD90 (77.3%) and negative for CD34 (2.57%) and CD45 (2.5%). ( E ) Alizarin Red S, Toluidine blue, and Oil Red O staining were used to detect osteogenic, chondrogenic, and adipogenic differentiation, respectively (scale bar, 100 μm). ( F ) SA-β-gal staining detected senescent cells (scale bar, 100 μm). ( G ) Western blotting analysis of Nanog, Sox2, and Oct4 protein expression in BMSCs from different groups. ( H ) Western blotting analysis of P53, P21, and P16 protein expression in BMSCs from different groups. ( I ) Representative immunofluorescence staining of γ-H2AX foci formation in BMSCs from different groups (scale bar, 25 μm). DAPI, 4′,6-diamidino-2-phenylindole. ( J ) Representative immunofluorescence staining of CD90 and Runx2 in BMSCs from the different groups (scale bar, 25 μm). ( K ) Representative immunofluorescence staining of CD90 and Opn in BMSCs from different groups (scale bar, 25 μm). ( L ) Western blotting analysis of Runx2 and Opn protein expression in BMSCs from different groups. ( M ) ALP staining of BMSCs from the different groups. ( N ) Alizarin Red S staining of BMSCs from different groups. n = 3 for each group. Error bars denote means ± SEM; ns, no significance; * P < 0.05, ** P < 0.01, and *** P < 0.001.

    Journal: Science Advances

    Article Title: Stem cell–homing biomimetic hydrogel promotes the repair of osteoporotic bone defects through osteogenic and angiogenic coupling

    doi: 10.1126/sciadv.adq6700

    Figure Lengend Snippet: ( A ) Micro-CT scan and 3D reconstruction results (scale bar, 1 mm); quantitative analysis of bone density, volume, and other parameters. BV/TV, bone tissue volume/total tissue; Tb.Th, trabecular thickness; Tb.N, trabecular number. ( B ) H&E staining images of different groups (scale bar, 1 mm and 125 μm). ( C ) Masson staining images of different groups (scale bar, 1 mm and 125 μm). ( D ) Flow cytometry showed BMSCs from osteoporotic rats were positive for CD44 (99.8%), CD29 (99.8%), CD73 (98.9%), and CD90 (77.3%) and negative for CD34 (2.57%) and CD45 (2.5%). ( E ) Alizarin Red S, Toluidine blue, and Oil Red O staining were used to detect osteogenic, chondrogenic, and adipogenic differentiation, respectively (scale bar, 100 μm). ( F ) SA-β-gal staining detected senescent cells (scale bar, 100 μm). ( G ) Western blotting analysis of Nanog, Sox2, and Oct4 protein expression in BMSCs from different groups. ( H ) Western blotting analysis of P53, P21, and P16 protein expression in BMSCs from different groups. ( I ) Representative immunofluorescence staining of γ-H2AX foci formation in BMSCs from different groups (scale bar, 25 μm). DAPI, 4′,6-diamidino-2-phenylindole. ( J ) Representative immunofluorescence staining of CD90 and Runx2 in BMSCs from the different groups (scale bar, 25 μm). ( K ) Representative immunofluorescence staining of CD90 and Opn in BMSCs from different groups (scale bar, 25 μm). ( L ) Western blotting analysis of Runx2 and Opn protein expression in BMSCs from different groups. ( M ) ALP staining of BMSCs from the different groups. ( N ) Alizarin Red S staining of BMSCs from different groups. n = 3 for each group. Error bars denote means ± SEM; ns, no significance; * P < 0.05, ** P < 0.01, and *** P < 0.001.

    Article Snippet: Following two washes in PBS, BMSCs were incubated for 6 hours in ALP staining buffer (Beyotime Institute of Biotechnology, China).

    Techniques: Micro-CT, Staining, Flow Cytometry, Western Blot, Expressing, Immunofluorescence

    ( A ) Immunofluorescence staining of F-actin in BMSCs cultured with hydrogels for 7 days (scale bar, 25 μm). ( B ) Live/dead assay of BMSCs cultured with hydrogels for 3 days (scale bar, 100 μm). Live cells appear green, and dead cells appear red. ( C ) Western blots analysis of P-eNOS and eNOS levels in BMSCs cultured with hydrogels for 7 days. ( D ) NO generation in BMSCs cultured with hydrogels for 7 days indicated by nitrite levels. ( E ) cGMP expression in BMSCs cultured with hydrogels for 7 days. ( F ) Western blots analysis of sGC and PKG levels in BMSCs cultured with hydrogels for 7 days. ( G ) Western blots analysis of Runx2 and Opn levels in BMSCs cultured with hydrogels for 7 days. ( H ) Representative of immunofluorescence staining of CD90 and Runx2 in BMSCs cultured with hydrogels for 7 days (scale bar, 25 μm). ( I ) Representative of immunofluorescence staining of CD90 and Opn in BMSCs cultured with hydrogels for 7 days (scale bar, 25 μm). ( J ) The ALP staining of BMSCs cultured with hydrogels for 7 days. ( K ) Alizarin Red S staining of BMSCs cultured with hydrogels for 7 days. ( L ) Schematic diagram of SA-MSNs@CM-Stiff activating NO/cGMP pathway to promote BMSC osteogenesis. n = 3 for each group. Error bars denote means ± SEM; * P < 0.05, ** P < 0.01, and *** P < 0.001.

    Journal: Science Advances

    Article Title: Stem cell–homing biomimetic hydrogel promotes the repair of osteoporotic bone defects through osteogenic and angiogenic coupling

    doi: 10.1126/sciadv.adq6700

    Figure Lengend Snippet: ( A ) Immunofluorescence staining of F-actin in BMSCs cultured with hydrogels for 7 days (scale bar, 25 μm). ( B ) Live/dead assay of BMSCs cultured with hydrogels for 3 days (scale bar, 100 μm). Live cells appear green, and dead cells appear red. ( C ) Western blots analysis of P-eNOS and eNOS levels in BMSCs cultured with hydrogels for 7 days. ( D ) NO generation in BMSCs cultured with hydrogels for 7 days indicated by nitrite levels. ( E ) cGMP expression in BMSCs cultured with hydrogels for 7 days. ( F ) Western blots analysis of sGC and PKG levels in BMSCs cultured with hydrogels for 7 days. ( G ) Western blots analysis of Runx2 and Opn levels in BMSCs cultured with hydrogels for 7 days. ( H ) Representative of immunofluorescence staining of CD90 and Runx2 in BMSCs cultured with hydrogels for 7 days (scale bar, 25 μm). ( I ) Representative of immunofluorescence staining of CD90 and Opn in BMSCs cultured with hydrogels for 7 days (scale bar, 25 μm). ( J ) The ALP staining of BMSCs cultured with hydrogels for 7 days. ( K ) Alizarin Red S staining of BMSCs cultured with hydrogels for 7 days. ( L ) Schematic diagram of SA-MSNs@CM-Stiff activating NO/cGMP pathway to promote BMSC osteogenesis. n = 3 for each group. Error bars denote means ± SEM; * P < 0.05, ** P < 0.01, and *** P < 0.001.

    Article Snippet: Following two washes in PBS, BMSCs were incubated for 6 hours in ALP staining buffer (Beyotime Institute of Biotechnology, China).

    Techniques: Immunofluorescence, Staining, Cell Culture, Live Dead Assay, Western Blot, Expressing

    MC3T3-E1 cells increased osteogenesis in the presence of BN. Notes: ( A ) Molecular structure of BN. ( B ) CCK-8 assays showed that BN did not significantly affect the growth of MC3T3-E1 cells at the concentrations used (0.1 μmol/L, 1 μmol/L, 10 μmol/L, 100 μmol/L, and 1,000 μmol/L) after treatment for 14 days, and BN at concentrations of 0.1–100 μM did not significantly affect cell growth. ( C ) The ALP activity assay showed that BN increased intracellular ALP expression during osteoblast differentiation in cultured MC3T3-E1 cells. ( D ) ALP staining indicates that treatment with 0.1–10 μM BN increased ALP activity in MC3T3-E1 cells in osteogenic differentiation medium for 14 days. ( E ) Micrograph of ALP staining indicates that the expression of ALP shows dose-dependent with 0.1–10 μM BN treatment. * P <0.05 versus group without BN; + P <0.05 versus group with 0.1 μM BN; # P <0.05 versus group with 1 μM BN. Columns represent the mean ± standard error from six wells per group ( B , C ). Abbreviations: ALP, alkaline phosphatase; BN, baicalein; CCK-8, Cell Counting Kit-8; OD, optical density.

    Journal: Drug Design, Development and Therapy

    Article Title: Regulation of bone formation by baicalein via the mTORC1 pathway

    doi: 10.2147/DDDT.S81578

    Figure Lengend Snippet: MC3T3-E1 cells increased osteogenesis in the presence of BN. Notes: ( A ) Molecular structure of BN. ( B ) CCK-8 assays showed that BN did not significantly affect the growth of MC3T3-E1 cells at the concentrations used (0.1 μmol/L, 1 μmol/L, 10 μmol/L, 100 μmol/L, and 1,000 μmol/L) after treatment for 14 days, and BN at concentrations of 0.1–100 μM did not significantly affect cell growth. ( C ) The ALP activity assay showed that BN increased intracellular ALP expression during osteoblast differentiation in cultured MC3T3-E1 cells. ( D ) ALP staining indicates that treatment with 0.1–10 μM BN increased ALP activity in MC3T3-E1 cells in osteogenic differentiation medium for 14 days. ( E ) Micrograph of ALP staining indicates that the expression of ALP shows dose-dependent with 0.1–10 μM BN treatment. * P <0.05 versus group without BN; + P <0.05 versus group with 0.1 μM BN; # P <0.05 versus group with 1 μM BN. Columns represent the mean ± standard error from six wells per group ( B , C ). Abbreviations: ALP, alkaline phosphatase; BN, baicalein; CCK-8, Cell Counting Kit-8; OD, optical density.

    Article Snippet: Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, washed, incubated with ALP staining buffer (NBT-BCIP, Sigma-Aldrich) at 37°C for 30 minutes, and washed with PBS to remove excess dye.

    Techniques: CCK-8 Assay, ALP Activity Assay, Expressing, Cell Culture, Staining, Activity Assay, Cell Counting