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    Thermo Fisher β glycerophosphate
    Osteoblastic characteristics of the OBN4 clone. ( a ) ALP activity of the OBN4 clone. ALP activities were analyzed between OBN4 and primary osteoblast (pOB) cells via stimulation with 50 μ M ascorbic acid, 10 m M <t>β-glycerophosphate</t> and 10 −7 M dexamethasone for 6 d. Photographs of ALP-positive cells in triplicate samples are presented (top panel), and ALP activities were quantified (bottom panel). ( b ) Mineralization of the OBN4 clone. The mineralization of cultured cells with 50 μ M ascorbic acid and 10 m M β-glycerophosphate for 21 d was measured using ARS staining. ARS-positive cells in triplicate samples were photographed (top panel), and ARS activities were quantified (bottom panel). ( c ) Expression of osteoblast markers in the OBN4 clone. The expression levels of osteocalcin (top panel) and osteopontin (bottom panel) were analyzed via real-time PCR analysis. ( d ) Expression of the Sca-1 lineage marker in the OBN4 clone. The expression of Sca-1 antigen was analyzed via flow cytometry with an anti-Sca-1 antibody (top panel). The percentages of the Sca-1 + cell population are summarized in the bottom panel. Mock, non-stained cells. ( e ) Comparison of the ability to support osteoclast differentiation in coculture assays. Cells were cocultured with bone marrow-derived HSCs in the presence of vitamin D3 and prostaglandin E2 for 6 d. Osteoclast formation was analyzed through TRAP staining (top panel). The number of TRAP-positive multinucleated osteoclasts (TRAP + MNCs (⩾three nuclei)) was counted (bottom panel). ( f ) RANKL expression in the OBN4 clone. The surface expression of RANKL was analyzed via flow cytometry with an anti-RANKL antibody (top panel). The percentages of the RANKL + cell population are summarized in the bottom panel. NS, not significant. * P
    β Glycerophosphate, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 59 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/β glycerophosphate/product/Thermo Fisher
    Average 93 stars, based on 59 article reviews
    Price from $9.99 to $1999.99
    β glycerophosphate - by Bioz Stars, 2020-09
    93/100 stars

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    1) Product Images from "Generation of an osteoblast-based artificial niche that supports in vitro B lymphopoiesis"

    Article Title: Generation of an osteoblast-based artificial niche that supports in vitro B lymphopoiesis

    Journal: Experimental & Molecular Medicine

    doi: 10.1038/emm.2017.189

    Osteoblastic characteristics of the OBN4 clone. ( a ) ALP activity of the OBN4 clone. ALP activities were analyzed between OBN4 and primary osteoblast (pOB) cells via stimulation with 50 μ M ascorbic acid, 10 m M β-glycerophosphate and 10 −7 M dexamethasone for 6 d. Photographs of ALP-positive cells in triplicate samples are presented (top panel), and ALP activities were quantified (bottom panel). ( b ) Mineralization of the OBN4 clone. The mineralization of cultured cells with 50 μ M ascorbic acid and 10 m M β-glycerophosphate for 21 d was measured using ARS staining. ARS-positive cells in triplicate samples were photographed (top panel), and ARS activities were quantified (bottom panel). ( c ) Expression of osteoblast markers in the OBN4 clone. The expression levels of osteocalcin (top panel) and osteopontin (bottom panel) were analyzed via real-time PCR analysis. ( d ) Expression of the Sca-1 lineage marker in the OBN4 clone. The expression of Sca-1 antigen was analyzed via flow cytometry with an anti-Sca-1 antibody (top panel). The percentages of the Sca-1 + cell population are summarized in the bottom panel. Mock, non-stained cells. ( e ) Comparison of the ability to support osteoclast differentiation in coculture assays. Cells were cocultured with bone marrow-derived HSCs in the presence of vitamin D3 and prostaglandin E2 for 6 d. Osteoclast formation was analyzed through TRAP staining (top panel). The number of TRAP-positive multinucleated osteoclasts (TRAP + MNCs (⩾three nuclei)) was counted (bottom panel). ( f ) RANKL expression in the OBN4 clone. The surface expression of RANKL was analyzed via flow cytometry with an anti-RANKL antibody (top panel). The percentages of the RANKL + cell population are summarized in the bottom panel. NS, not significant. * P
    Figure Legend Snippet: Osteoblastic characteristics of the OBN4 clone. ( a ) ALP activity of the OBN4 clone. ALP activities were analyzed between OBN4 and primary osteoblast (pOB) cells via stimulation with 50 μ M ascorbic acid, 10 m M β-glycerophosphate and 10 −7 M dexamethasone for 6 d. Photographs of ALP-positive cells in triplicate samples are presented (top panel), and ALP activities were quantified (bottom panel). ( b ) Mineralization of the OBN4 clone. The mineralization of cultured cells with 50 μ M ascorbic acid and 10 m M β-glycerophosphate for 21 d was measured using ARS staining. ARS-positive cells in triplicate samples were photographed (top panel), and ARS activities were quantified (bottom panel). ( c ) Expression of osteoblast markers in the OBN4 clone. The expression levels of osteocalcin (top panel) and osteopontin (bottom panel) were analyzed via real-time PCR analysis. ( d ) Expression of the Sca-1 lineage marker in the OBN4 clone. The expression of Sca-1 antigen was analyzed via flow cytometry with an anti-Sca-1 antibody (top panel). The percentages of the Sca-1 + cell population are summarized in the bottom panel. Mock, non-stained cells. ( e ) Comparison of the ability to support osteoclast differentiation in coculture assays. Cells were cocultured with bone marrow-derived HSCs in the presence of vitamin D3 and prostaglandin E2 for 6 d. Osteoclast formation was analyzed through TRAP staining (top panel). The number of TRAP-positive multinucleated osteoclasts (TRAP + MNCs (⩾three nuclei)) was counted (bottom panel). ( f ) RANKL expression in the OBN4 clone. The surface expression of RANKL was analyzed via flow cytometry with an anti-RANKL antibody (top panel). The percentages of the RANKL + cell population are summarized in the bottom panel. NS, not significant. * P

    Techniques Used: ALP Assay, Activity Assay, Cell Culture, Staining, Expressing, Real-time Polymerase Chain Reaction, Marker, Flow Cytometry, Cytometry, Derivative Assay

    Development of the OBN4 clone. ( a ) Schematic diagram of the SV40 Tag expression plasmid. FLAG-epitope (hatched box); LTR, retroviral long terminal repeat; Puro R , puromycin resistant cassette. ( b ) Expression of FLAG-tagged SV40 Tag in OBN cells. The expression of SV40 Tag was analyzed by western blotting with an anti-FLAG antibody. β-Actin was used as a loading control. pOB, primary osteoblast. ( c ) Comparison of ALP activities in OBN clones. The ALP activity levels of single OBN clones were analyzed through stimulation with 50 μ M ascorbic acid, 10 m M β-glycerophosphate and 10 −7 M dexamethasone for 6 d. * P
    Figure Legend Snippet: Development of the OBN4 clone. ( a ) Schematic diagram of the SV40 Tag expression plasmid. FLAG-epitope (hatched box); LTR, retroviral long terminal repeat; Puro R , puromycin resistant cassette. ( b ) Expression of FLAG-tagged SV40 Tag in OBN cells. The expression of SV40 Tag was analyzed by western blotting with an anti-FLAG antibody. β-Actin was used as a loading control. pOB, primary osteoblast. ( c ) Comparison of ALP activities in OBN clones. The ALP activity levels of single OBN clones were analyzed through stimulation with 50 μ M ascorbic acid, 10 m M β-glycerophosphate and 10 −7 M dexamethasone for 6 d. * P

    Techniques Used: Expressing, Plasmid Preparation, FLAG-tag, Western Blot, ALP Assay, Clone Assay, Activity Assay

    2) Product Images from "Generation of an osteoblast-based artificial niche that supports in vitro B lymphopoiesis"

    Article Title: Generation of an osteoblast-based artificial niche that supports in vitro B lymphopoiesis

    Journal: Experimental & Molecular Medicine

    doi: 10.1038/emm.2017.189

    Osteoblastic characteristics of the OBN4 clone. ( a ) ALP activity of the OBN4 clone. ALP activities were analyzed between OBN4 and primary osteoblast (pOB) cells via stimulation with 50 μ M ascorbic acid, 10 m M β-glycerophosphate and 10 −7 M dexamethasone for 6 d. Photographs of ALP-positive cells in triplicate samples are presented (top panel), and ALP activities were quantified (bottom panel). ( b ) Mineralization of the OBN4 clone. The mineralization of cultured cells with 50 μ M ascorbic acid and 10 m M β-glycerophosphate for 21 d was measured using ARS staining. ARS-positive cells in triplicate samples were photographed (top panel), and ARS activities were quantified (bottom panel). ( c ) Expression of osteoblast markers in the OBN4 clone. The expression levels of osteocalcin (top panel) and osteopontin (bottom panel) were analyzed via real-time PCR analysis. ( d ) Expression of the Sca-1 lineage marker in the OBN4 clone. The expression of Sca-1 antigen was analyzed via flow cytometry with an anti-Sca-1 antibody (top panel). The percentages of the Sca-1 + cell population are summarized in the bottom panel. Mock, non-stained cells. ( e ) Comparison of the ability to support osteoclast differentiation in coculture assays. Cells were cocultured with bone marrow-derived HSCs in the presence of vitamin D3 and prostaglandin E2 for 6 d. Osteoclast formation was analyzed through TRAP staining (top panel). The number of TRAP-positive multinucleated osteoclasts (TRAP + MNCs (⩾three nuclei)) was counted (bottom panel). ( f ) RANKL expression in the OBN4 clone. The surface expression of RANKL was analyzed via flow cytometry with an anti-RANKL antibody (top panel). The percentages of the RANKL + cell population are summarized in the bottom panel. NS, not significant. * P
    Figure Legend Snippet: Osteoblastic characteristics of the OBN4 clone. ( a ) ALP activity of the OBN4 clone. ALP activities were analyzed between OBN4 and primary osteoblast (pOB) cells via stimulation with 50 μ M ascorbic acid, 10 m M β-glycerophosphate and 10 −7 M dexamethasone for 6 d. Photographs of ALP-positive cells in triplicate samples are presented (top panel), and ALP activities were quantified (bottom panel). ( b ) Mineralization of the OBN4 clone. The mineralization of cultured cells with 50 μ M ascorbic acid and 10 m M β-glycerophosphate for 21 d was measured using ARS staining. ARS-positive cells in triplicate samples were photographed (top panel), and ARS activities were quantified (bottom panel). ( c ) Expression of osteoblast markers in the OBN4 clone. The expression levels of osteocalcin (top panel) and osteopontin (bottom panel) were analyzed via real-time PCR analysis. ( d ) Expression of the Sca-1 lineage marker in the OBN4 clone. The expression of Sca-1 antigen was analyzed via flow cytometry with an anti-Sca-1 antibody (top panel). The percentages of the Sca-1 + cell population are summarized in the bottom panel. Mock, non-stained cells. ( e ) Comparison of the ability to support osteoclast differentiation in coculture assays. Cells were cocultured with bone marrow-derived HSCs in the presence of vitamin D3 and prostaglandin E2 for 6 d. Osteoclast formation was analyzed through TRAP staining (top panel). The number of TRAP-positive multinucleated osteoclasts (TRAP + MNCs (⩾three nuclei)) was counted (bottom panel). ( f ) RANKL expression in the OBN4 clone. The surface expression of RANKL was analyzed via flow cytometry with an anti-RANKL antibody (top panel). The percentages of the RANKL + cell population are summarized in the bottom panel. NS, not significant. * P

    Techniques Used: ALP Assay, Activity Assay, Cell Culture, Staining, Expressing, Real-time Polymerase Chain Reaction, Marker, Flow Cytometry, Cytometry, Derivative Assay

    Development of the OBN4 clone. ( a ) Schematic diagram of the SV40 Tag expression plasmid. FLAG-epitope (hatched box); LTR, retroviral long terminal repeat; Puro R , puromycin resistant cassette. ( b ) Expression of FLAG-tagged SV40 Tag in OBN cells. The expression of SV40 Tag was analyzed by western blotting with an anti-FLAG antibody. β-Actin was used as a loading control. pOB, primary osteoblast. ( c ) Comparison of ALP activities in OBN clones. The ALP activity levels of single OBN clones were analyzed through stimulation with 50 μ M ascorbic acid, 10 m M β-glycerophosphate and 10 −7 M dexamethasone for 6 d. * P
    Figure Legend Snippet: Development of the OBN4 clone. ( a ) Schematic diagram of the SV40 Tag expression plasmid. FLAG-epitope (hatched box); LTR, retroviral long terminal repeat; Puro R , puromycin resistant cassette. ( b ) Expression of FLAG-tagged SV40 Tag in OBN cells. The expression of SV40 Tag was analyzed by western blotting with an anti-FLAG antibody. β-Actin was used as a loading control. pOB, primary osteoblast. ( c ) Comparison of ALP activities in OBN clones. The ALP activity levels of single OBN clones were analyzed through stimulation with 50 μ M ascorbic acid, 10 m M β-glycerophosphate and 10 −7 M dexamethasone for 6 d. * P

    Techniques Used: Expressing, Plasmid Preparation, FLAG-tag, Western Blot, ALP Assay, Clone Assay, Activity Assay

    Related Articles

    Transfection:

    Article Title: Generation of an osteoblast-based artificial niche that supports in vitro B lymphopoiesis
    Article Snippet: .. The collected cells were differentiated into primary osteoblasts by culture in α-MEM including 10% FBS and supplemented with 50 μM ascorbic acid, 10 mM β-glycerophosphate, 100 ngml−1 BMP-2 and 10−7 M dexamethasone for 5 d. The primary osteoblasts were further activated by treatment with 50 μM ascorbic acid, 10 mM β-glycerophosphate and 10−7 M PTH for 3 d. To prepare a retroviral supernatant harboring the SV40 Tag expression cassette, PlatE cells were transfected with retroviral SV40 Tag expression plasmid DNA using the TurboFect transfection reagent (Fermentas, Hanover, MD) according to the manufacturer's instructions. .. Subsequently, the PTH-stimulated primary osteoblasts were infected with the retroviral supernatant harboring the SV40 Tag expression cassette with polybrene (8 μgml−1 ) for 6 h. The retrovirally transduced cells were selected with puromycin (2 μgml−1 ) for 14 d to generate a stable cell line.

    Protease Inhibitor:

    Article Title: γ-Oryzanol Enhances Adipocyte Differentiation and Glucose Uptake
    Article Snippet: .. The cells were lysed with a lysis buffer containing 40 mM HEPES (pH 7.4), 120 mM sodium chloride (NaCl), 1 mM ethylenediaminetetraacetic acid (EDTA), 50 mM sodium fluoride (NaF), 1.5 mM sodium orthovanadate (Na3 VO4 ), 10 mM β-glycerophosphate, and 1% Triton X-100 supplemented with EDTA-free phosphatase and protease inhibitor cocktail (#78441, Thermo Fisher Scientific Inc., Rockford, IL, USA). .. After centrifugation at 14,000× g for 20 min at 4 °C, the supernatants were boiled in sodium dodecyl sulfate (SDS)-loading buffer.

    Cell Culture:

    Article Title: Silk Biomaterials-Mediated miRNA Functionalized Orthopedic Devices
    Article Snippet: .. To assess early osteoinductivity, hMSCs were cultured with osteogenic medium containing 10 nM dexamethasone, 10 mM β-glycerophosphate, and 50 μg/mL ascorbic acid for 2 weeks and stained with commercially available Vector Blue (Invitrogen, Carlsbad, CA). .. For ALP staining, cells were fixed in 4% paraformaldehyde for 15 min at room temperature and washed with PBS.

    Incubation:

    Article Title: Maintenance of the DNA-Damage Checkpoint Requires DNA-Damage-Induced Mediator Protein Oligomerization
    Article Snippet: .. After washed, anti-HA IPs were incubated in 150 μl of TEV buffer (10 mM Tris-HCl [pH 8.0], 0.5 mM EDTA, 1 mM DTT, 0.1% NP-40, 150 mM NaCl, 2 mM NaF, 5 mM β-glycerophosphate) with AcTEV protease (Invitrogen, 20 units/IP) at 30°C for 1.5 hr. .. Supernatants that contain N-SCD were collected and incubated with 0.1 μg GST-BRCT proteins bound to 10 μl of glutathione Sepharose 4B (GE Healthcare) at 4°C for 2 hr.

    Expressing:

    Article Title: Generation of an osteoblast-based artificial niche that supports in vitro B lymphopoiesis
    Article Snippet: .. The collected cells were differentiated into primary osteoblasts by culture in α-MEM including 10% FBS and supplemented with 50 μM ascorbic acid, 10 mM β-glycerophosphate, 100 ngml−1 BMP-2 and 10−7 M dexamethasone for 5 d. The primary osteoblasts were further activated by treatment with 50 μM ascorbic acid, 10 mM β-glycerophosphate and 10−7 M PTH for 3 d. To prepare a retroviral supernatant harboring the SV40 Tag expression cassette, PlatE cells were transfected with retroviral SV40 Tag expression plasmid DNA using the TurboFect transfection reagent (Fermentas, Hanover, MD) according to the manufacturer's instructions. .. Subsequently, the PTH-stimulated primary osteoblasts were infected with the retroviral supernatant harboring the SV40 Tag expression cassette with polybrene (8 μgml−1 ) for 6 h. The retrovirally transduced cells were selected with puromycin (2 μgml−1 ) for 14 d to generate a stable cell line.

    Staining:

    Article Title: Silk Biomaterials-Mediated miRNA Functionalized Orthopedic Devices
    Article Snippet: .. To assess early osteoinductivity, hMSCs were cultured with osteogenic medium containing 10 nM dexamethasone, 10 mM β-glycerophosphate, and 50 μg/mL ascorbic acid for 2 weeks and stained with commercially available Vector Blue (Invitrogen, Carlsbad, CA). .. For ALP staining, cells were fixed in 4% paraformaldehyde for 15 min at room temperature and washed with PBS.

    Lysis:

    Article Title: MicroRNA-126 overexpression rescues diabetes-induced impairment in efferocytosis of apoptotic cardiomyocytes
    Article Snippet: .. Briefly, cells were homogenized in lysis buffer (Cell Signaling Technology, MA, USA) containing 20 mmol/l Tris-HCl [pH 7.5], 150 mmol/l NaCl, 2.5 mmol/l sodium pyrophosphate, 1 mmol/l β-glycerophosphate, 1 mmol/l sodium orthovanadate, 1 μg/ml leupeptin, 1 mmol/l ethylenediaminetetraacetic acid [EDTA], 1 mmol/l ethylene glycol tetraacetic acid [EGTA], 1% Triton X-100, protease and phosphatase inhibitors (Thermo Fisher Scientific). .. Immunoprecipitation was performed by incubating cell lysates with 1 μg of MERTK antibody targeted to the intracellular C-terminus domain (Abcam, Cat # ab95925) or IgG control antibody for overnight (12 h) with gentle rotation.

    Article Title: γ-Oryzanol Enhances Adipocyte Differentiation and Glucose Uptake
    Article Snippet: .. The cells were lysed with a lysis buffer containing 40 mM HEPES (pH 7.4), 120 mM sodium chloride (NaCl), 1 mM ethylenediaminetetraacetic acid (EDTA), 50 mM sodium fluoride (NaF), 1.5 mM sodium orthovanadate (Na3 VO4 ), 10 mM β-glycerophosphate, and 1% Triton X-100 supplemented with EDTA-free phosphatase and protease inhibitor cocktail (#78441, Thermo Fisher Scientific Inc., Rockford, IL, USA). .. After centrifugation at 14,000× g for 20 min at 4 °C, the supernatants were boiled in sodium dodecyl sulfate (SDS)-loading buffer.

    Plasmid Preparation:

    Article Title: Generation of an osteoblast-based artificial niche that supports in vitro B lymphopoiesis
    Article Snippet: .. The collected cells were differentiated into primary osteoblasts by culture in α-MEM including 10% FBS and supplemented with 50 μM ascorbic acid, 10 mM β-glycerophosphate, 100 ngml−1 BMP-2 and 10−7 M dexamethasone for 5 d. The primary osteoblasts were further activated by treatment with 50 μM ascorbic acid, 10 mM β-glycerophosphate and 10−7 M PTH for 3 d. To prepare a retroviral supernatant harboring the SV40 Tag expression cassette, PlatE cells were transfected with retroviral SV40 Tag expression plasmid DNA using the TurboFect transfection reagent (Fermentas, Hanover, MD) according to the manufacturer's instructions. .. Subsequently, the PTH-stimulated primary osteoblasts were infected with the retroviral supernatant harboring the SV40 Tag expression cassette with polybrene (8 μgml−1 ) for 6 h. The retrovirally transduced cells were selected with puromycin (2 μgml−1 ) for 14 d to generate a stable cell line.

    Article Title: Silk Biomaterials-Mediated miRNA Functionalized Orthopedic Devices
    Article Snippet: .. To assess early osteoinductivity, hMSCs were cultured with osteogenic medium containing 10 nM dexamethasone, 10 mM β-glycerophosphate, and 50 μg/mL ascorbic acid for 2 weeks and stained with commercially available Vector Blue (Invitrogen, Carlsbad, CA). .. For ALP staining, cells were fixed in 4% paraformaldehyde for 15 min at room temperature and washed with PBS.

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    Thermo Fisher β glycerophosphate
    β Glycerophosphate, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 59 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/β glycerophosphate/product/Thermo Fisher
    Average 93 stars, based on 59 article reviews
    Price from $9.99 to $1999.99
    β glycerophosphate - by Bioz Stars, 2020-09
    93/100 stars
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