NIH3T3-Light2 cells细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心
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NIH3T3-Light2 cells细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心
NIH3T3 cells， NIH3T3-Light2 cells
Derived from NIH/3T3 cell line in 1999.
MoreLess NIH/3T3 cells were co-transfected with GLI-responsive Firefly luciferase reporter from H. Sasaki et al. (Development 1997 Vol. 7: 1313-22), and pSV-Neo from Daniel Nathans. A stable clonal cell line was selected using G418. The resulting cell line has been transfected with pRL-TK constituitive Renilla-luciferase expression vector (Promega), and pVgRXR vector (Invitrogen) encoding the ecdysone receptor and a Zeocin resistance marker. Zeocin selection and cell cloning was then used to generate Shh Light II cells. Useful as a cell-based tool for the quantitative assay of biologically active hedgehog proteins. These cells respond to the presence of hedgehog protein by producing light-generating enzyme that can be measured with a luminometer.Assay Conditions:Cells cultured to maximal density should be incubated for 30-48 h in DMEM supplemented with 0.5% calf serum and 5 mM HEPES buffer (pH 7.4) and the tested sample. Please note that cultures of NIH/3T3 cells that are commonly referred to as confluent take approximately 1 to 2 days to reach maximal cell density (full contact inhibition of growth). Unless transfection efficiency is extremely high (30-100%), this cell line is not suitable for transient transfection-based assays using constructs whose effects are cell autonomous. Parental NIH/3T3 cells can be used instead, and the tested vector co-transfected with Hh-sensitive reporters.
Organism Mus musculus, mouse
Tissue embryo
Product Format frozen 1 mL per vial; 1 x 106 cells/mL.
Morphology fibroblast
Culture Properties adherent
Biosafety Level 2
Age embryo
Applications
Parental NIH/3T3 cells can be used instead, and the tested vector co-transfected with Hh-sensitive reporters.
Zeocin selection and cell cloning was then used to generate Shh Light II cells.
Useful as a cell-based tool for the quantitative assay of biologically active hedgehog proteins.
Unless transfection efficiency is extremely high (30-100%), this cell line is not suitable for transient transfection-based assays using constructs whose effects are cell autonomous.
Derived from NIH/3T3 cell line in 1999. NIH/3T3 cells were co-transfected with GLI-responsive Firefly luciferase reporter from H. Sasaki et al.
These cells respond to the presence of hedgehog protein by producing light-generating enzyme that can be measured with a luminometer.
Derivation
Derived from NIH/3T3 cell line in 1999. NIH/3T3 cells were co-transfected with GLI-responsive Firefly luciferase reporter from H. Sasaki et al. (Development 1997 Vol. 7: 1313-22), and pSV-Neo from Daniel Nathans. A stable clonal cell line was selected using G418. The resulting cell line has been transfected with pRL-TK constituitive Renilla-luciferase expression vector (Promega), and pVgRXR vector (Invitrogen) encoding the ecdysone receptor and a Zeocin resistance marker. Zeocin selection and cell cloning was then used to generate Shh Light II cells. Useful as a cell-based tool for the quantitative assay of biologically active hedgehog proteins. These cells respond to the presence of hedgehog protein by producing light-generating enzyme that can be measured with a luminometer.Assay Conditions:Cells cultured to maximal density should be incubated for 30-48 h in DMEM supplemented with 0.5% calf serum and 5 mM HEPES buffer (pH 7.4) and the tested sample. Please note that cultures of NIH/3T3 cells that are commonly referred to as confluent take approximately 1 to 2 days to reach maximal cell density (full contact inhibition of growth). Unless transfection efficiency is extremely high (30-100%), this cell line is not suitable for transient transfection-based assays using constructs whose effects are cell autonomous. Parental NIH/3T3 cells can be used instead, and the tested vector co-transfected with Hh-sensitive reporters.
Comments
Derived from NIH/3T3 cell line in 1999. NIH/3T3 cells were co-transfected with GLI-responsive Firefly luciferase reporter from H. Sasaki et al. (Development 1997 Vol. 7: 1313-22), and pSV-Neo from Daniel Nathans. A stable clonal cell line was selected using G418. The resulting cell line has been transfected with pRL-TK constituitive Renilla-luciferase expression vector (Promega), and pVgRXR vector (Invitrogen) encoding the ecdysone receptor and a Zeocin resistance marker. Zeocin selection and cell cloning was then used to generate Shh Light II cells. Useful as a cell-based tool for the quantitative assay of biologically active hedgehog proteins. These cells respond to the presence of hedgehog protein by producing light-generating enzyme that can be measured with a luminometer.
Assay Conditions:
Cells cultured to maximal density should be incubated for 30-48 h in DMEM supplemented with 0.5% calf serum and 5 mM HEPES buffer (pH 7.4) and the tested sample. Please note that cultures of NIH/3T3 cells that are commonly referred to as confluent take approximately 1 to 2 days to reach maximal cell density (full contact inhibition of growth). Unless transfection efficiency is extremely high (30-100%), this cell line is not suitable for transient transfection-based assays using constructs whose effects are cell autonomous. Parental NIH/3T3 cells can be used instead, and the tested vector co-transfected with Hh-sensitive reporters.
Culture Method
Complete Growth Medium Dulbecco's modified Eagle's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose supplemented with 0.4 mg/ml G-418, 0.15 mg/ml Zeocin (Invitrogen, Cat. No. R25001) and 10% bovine calf serum.
Subculturing
Protocol: Subculture using standard trypsinization procedures. Centrifuge to remove trypsin.
Subcultivation Ratio: A subcultivation ratio of 1:4 every three days is recommended
Medium Renewal: 2 to 3 times per week
Cryopreservation
Freeze medium: Complete growth medium, 92%; DMSO, 8%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Temperature: 37.0°C
Specifications
Volume 1 mL per vial; 1 x 106 cells/mL.
History
Name of Depositor PA Beachy
Year of Origin 1989
References
Taipale J, et al. Effects of oncogenic mutations in Smoothened and Patched can be reversed by cyclopamine. Nature 406: 1005-1009, 2000. PubMed: 10984056
Chen JK, et al. Small molecule modulation of Smoothened activity. Proc. Natl. Acad. Sci. USA : 14071-14076, 2002. PubMed: 12391318
Sasaki H, et al. A binding site for Gli proteins is essential for HNF-3beta floor plate enhancer activity in transgenics and can respond to Shh in vitro. Development 124: 1313-1322, 1997. PubMed: 9118802
【Supplier来源】BioVector NTCC Inc.
【Website网址】 http://www.biovector.net
NIH3T3 cells， NIH3T3-Light2 cells
Derived from NIH/3T3 cell line in 1999.
MoreLess NIH/3T3 cells were co-transfected with GLI-responsive Firefly luciferase reporter from H. Sasaki et al. (Development 1997 Vol. 7: 1313-22), and pSV-Neo from Daniel Nathans. A stable clonal cell line was selected using G418. The resulting cell line has been transfected with pRL-TK constituitive Renilla-luciferase expression vector (Promega), and pVgRXR vector (Invitrogen) encoding the ecdysone receptor and a Zeocin resistance marker. Zeocin selection and cell cloning was then used to generate Shh Light II cells. Useful as a cell-based tool for the quantitative assay of biologically active hedgehog proteins. These cells respond to the presence of hedgehog protein by producing light-generating enzyme that can be measured with a luminometer.Assay Conditions:Cells cultured to maximal density should be incubated for 30-48 h in DMEM supplemented with 0.5% calf serum and 5 mM HEPES buffer (pH 7.4) and the tested sample. Please note that cultures of NIH/3T3 cells that are commonly referred to as confluent take approximately 1 to 2 days to reach maximal cell density (full contact inhibition of growth). Unless transfection efficiency is extremely high (30-100%), this cell line is not suitable for transient transfection-based assays using constructs whose effects are cell autonomous. Parental NIH/3T3 cells can be used instead, and the tested vector co-transfected with Hh-sensitive reporters.
Organism Mus musculus, mouse
Tissue embryo
Product Format frozen 1 mL per vial; 1 x 106 cells/mL.
Morphology fibroblast
Culture Properties adherent
Biosafety Level 2
Age embryo
Applications
Parental NIH/3T3 cells can be used instead, and the tested vector co-transfected with Hh-sensitive reporters.
Zeocin selection and cell cloning was then used to generate Shh Light II cells.
Useful as a cell-based tool for the quantitative assay of biologically active hedgehog proteins.
Unless transfection efficiency is extremely high (30-100%), this cell line is not suitable for transient transfection-based assays using constructs whose effects are cell autonomous.
Derived from NIH/3T3 cell line in 1999. NIH/3T3 cells were co-transfected with GLI-responsive Firefly luciferase reporter from H. Sasaki et al.
These cells respond to the presence of hedgehog protein by producing light-generating enzyme that can be measured with a luminometer.
Derivation
Derived from NIH/3T3 cell line in 1999. NIH/3T3 cells were co-transfected with GLI-responsive Firefly luciferase reporter from H. Sasaki et al. (Development 1997 Vol. 7: 1313-22), and pSV-Neo from Daniel Nathans. A stable clonal cell line was selected using G418. The resulting cell line has been transfected with pRL-TK constituitive Renilla-luciferase expression vector (Promega), and pVgRXR vector (Invitrogen) encoding the ecdysone receptor and a Zeocin resistance marker. Zeocin selection and cell cloning was then used to generate Shh Light II cells. Useful as a cell-based tool for the quantitative assay of biologically active hedgehog proteins. These cells respond to the presence of hedgehog protein by producing light-generating enzyme that can be measured with a luminometer.Assay Conditions:Cells cultured to maximal density should be incubated for 30-48 h in DMEM supplemented with 0.5% calf serum and 5 mM HEPES buffer (pH 7.4) and the tested sample. Please note that cultures of NIH/3T3 cells that are commonly referred to as confluent take approximately 1 to 2 days to reach maximal cell density (full contact inhibition of growth). Unless transfection efficiency is extremely high (30-100%), this cell line is not suitable for transient transfection-based assays using constructs whose effects are cell autonomous. Parental NIH/3T3 cells can be used instead, and the tested vector co-transfected with Hh-sensitive reporters.
Comments
Derived from NIH/3T3 cell line in 1999. NIH/3T3 cells were co-transfected with GLI-responsive Firefly luciferase reporter from H. Sasaki et al. (Development 1997 Vol. 7: 1313-22), and pSV-Neo from Daniel Nathans. A stable clonal cell line was selected using G418. The resulting cell line has been transfected with pRL-TK constituitive Renilla-luciferase expression vector (Promega), and pVgRXR vector (Invitrogen) encoding the ecdysone receptor and a Zeocin resistance marker. Zeocin selection and cell cloning was then used to generate Shh Light II cells. Useful as a cell-based tool for the quantitative assay of biologically active hedgehog proteins. These cells respond to the presence of hedgehog protein by producing light-generating enzyme that can be measured with a luminometer.
Assay Conditions:
Cells cultured to maximal density should be incubated for 30-48 h in DMEM supplemented with 0.5% calf serum and 5 mM HEPES buffer (pH 7.4) and the tested sample. Please note that cultures of NIH/3T3 cells that are commonly referred to as confluent take approximately 1 to 2 days to reach maximal cell density (full contact inhibition of growth). Unless transfection efficiency is extremely high (30-100%), this cell line is not suitable for transient transfection-based assays using constructs whose effects are cell autonomous. Parental NIH/3T3 cells can be used instead, and the tested vector co-transfected with Hh-sensitive reporters.
Culture Method
Complete Growth Medium Dulbecco's modified Eagle's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose supplemented with 0.4 mg/ml G-418, 0.15 mg/ml Zeocin (Invitrogen, Cat. No. R25001) and 10% bovine calf serum.
Subculturing
Protocol: Subculture using standard trypsinization procedures. Centrifuge to remove trypsin.
Subcultivation Ratio: A subcultivation ratio of 1:4 every three days is recommended
Medium Renewal: 2 to 3 times per week
Cryopreservation
Freeze medium: Complete growth medium, 92%; DMSO, 8%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Temperature: 37.0°C
Specifications
Volume 1 mL per vial; 1 x 106 cells/mL.
History
Name of Depositor PA Beachy
Year of Origin 1989
References
Taipale J, et al. Effects of oncogenic mutations in Smoothened and Patched can be reversed by cyclopamine. Nature 406: 1005-1009, 2000. PubMed: 10984056
Chen JK, et al. Small molecule modulation of Smoothened activity. Proc. Natl. Acad. Sci. USA : 14071-14076, 2002. PubMed: 12391318
Sasaki H, et al. A binding site for Gli proteins is essential for HNF-3beta floor plate enhancer activity in transgenics and can respond to Shh in vitro. Development 124: 1313-1322, 1997. PubMed: 9118802
【Supplier来源】BioVector NTCC Inc.
【Website网址】 http://www.biovector.net
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