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PA317 LXSN 16E7
PA317 LXSN 16E7
规格:
货期:
编号:TS211140
品牌:Testobio
产品名称: PA317 LXSN 16E7
商品货号: TS211140
Organism: Mus musculus, mouse
Tissue: embryo
Cell Type: Retinoblastoma,keratinocyte
Product Format: frozen
Morphology: fibroblast
Culture Properties: adherent
Biosafety Level: 2 Cells contain human papilloma viral DNA sequences

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Disease: Papilloma,Retinoblastoma
Age: embryo
Strain: NIH/Swiss
Applications:
Virions produced from the transfected Psi-2 cells were used to infect the amphotropic packaging line PA317, and infected cells were selected in medium containing G418.
The pLXSN16E7 vector contains the human papilloma virus (HPV) type 16 E7 gene under control of the Moloney murine leukemia virus (MoMuLV) promoter- enhancer sequences.
The vector also contains a gene controlling resistance to neomycin transcribed from the SV40 promoter.
This line produces the amphotropic retrovirus LXSN16E6 which encodes the HPV16 E7 open reading frame, and which can be used to stably infect many cell types.
The resulting cells constitutively express the E6 protein of HPV16 which binds to and inactivates the retinoblastoma (Rb) gene.
The virus can be used to immortalize human keratinocytes.
Storage Conditions: liquid nitrogen vapor phase
Derivation:
PA317 LXSN 16E7 is a packaging cell line developed by transfection of the retrovirus vector pLXSN16E7 into the Psi-2 ecotropic packaging cell line.
Virions produced from the transfected Psi-2 cells were used to infect the amphotropic packaging line PA317, and infected cells were selected in medium containing G418.
Comments:
PA317 LXSN 16E7 is a packaging cell line developed by transfection of the retrovirus vector pLXSN16E7 into the Psi-2 ecotropic packaging cell line.
Virions produced from the transfected Psi-2 cells were used to infect the amphotropic packaging line PA317, and infected cells were selected in medium containing G418.
The pLXSN16E7 vector contains the human papilloma virus (HPV) type 16 E7 gene under control of the Moloney murine leukemia virus (MoMuLV) promoter- enhancer sequences.
The vector also contains a gene controlling resistance to neomycin transcribed from the SV40 promoter.
This line produces the amphotropic retrovirus LXSN16E6 which encodes the HPV16 E7 open reading frame, and which can be used to stably infect many cell types.
The resulting cells constitutively express the E6 protein of HPV16 which binds to and inactivates the retinoblastoma (Rb) gene.
The virus can be used to immortalize human keratinocytes.
Complete Growth Medium: The base medium for this cell line is ATCC-formulated Dulbeccos Modified Eagles Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Subculturing: Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
    Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C.

Subcultivation Ratio: 1:6 to 1:12
Medium Renewal: Every 2 to 3 days

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Cryopreservation:

Complete growth medium described above supplemented with 5% (v/v) DMSO.xa0 Cell culture tested DMSO is available as ATCC Catalog No. 4-X.

Culture Conditions:
Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5%
Name of Depositor: DA Galloway
Deposited As: Mus musculus
References:

Halbert CL, et al. The E7 gene of human papillomavirus type 16 is sufficient for immortalization of human epithelial cells. J. Virol. 65: 473-478, 1991. PubMed: 1845902

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

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PA317 LXSN 16E7

  • 货号: TS211140
  • 好评
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  • 品牌 : TESTOBIO
产品名称: PA317 LXSN 16E7
商品货号: TS211140
Organism: Mus musculus, mouse
Tissue: embryo
Cell Type: Retinoblastoma,keratinocyte
Product Format: frozen
Morphology: fibroblast
Culture Properties: adherent
Biosafety Level: 2 Cells contain human papilloma viral DNA sequences

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Disease: Papilloma,Retinoblastoma
Age: embryo
Strain: NIH/Swiss
Applications:
Virions produced from the transfected Psi-2 cells were used to infect the amphotropic packaging line PA317, and infected cells were selected in medium containing G418.
The pLXSN16E7 vector contains the human papilloma virus (HPV) type 16 E7 gene under control of the Moloney murine leukemia virus (MoMuLV) promoter- enhancer sequences.
The vector also contains a gene controlling resistance to neomycin transcribed from the SV40 promoter.
This line produces the amphotropic retrovirus LXSN16E6 which encodes the HPV16 E7 open reading frame, and which can be used to stably infect many cell types.
The resulting cells constitutively express the E6 protein of HPV16 which binds to and inactivates the retinoblastoma (Rb) gene.
The virus can be used to immortalize human keratinocytes.
Storage Conditions: liquid nitrogen vapor phase
Derivation:
PA317 LXSN 16E7 is a packaging cell line developed by transfection of the retrovirus vector pLXSN16E7 into the Psi-2 ecotropic packaging cell line.
Virions produced from the transfected Psi-2 cells were used to infect the amphotropic packaging line PA317, and infected cells were selected in medium containing G418.
Comments:
PA317 LXSN 16E7 is a packaging cell line developed by transfection of the retrovirus vector pLXSN16E7 into the Psi-2 ecotropic packaging cell line.
Virions produced from the transfected Psi-2 cells were used to infect the amphotropic packaging line PA317, and infected cells were selected in medium containing G418.
The pLXSN16E7 vector contains the human papilloma virus (HPV) type 16 E7 gene under control of the Moloney murine leukemia virus (MoMuLV) promoter- enhancer sequences.
The vector also contains a gene controlling resistance to neomycin transcribed from the SV40 promoter.
This line produces the amphotropic retrovirus LXSN16E6 which encodes the HPV16 E7 open reading frame, and which can be used to stably infect many cell types.
The resulting cells constitutively express the E6 protein of HPV16 which binds to and inactivates the retinoblastoma (Rb) gene.
The virus can be used to immortalize human keratinocytes.
Complete Growth Medium: The base medium for this cell line is ATCC-formulated Dulbeccos Modified Eagles Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Subculturing: Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
    Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C.

Subcultivation Ratio: 1:6 to 1:12
Medium Renewal: Every 2 to 3 days

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Cryopreservation:

Complete growth medium described above supplemented with 5% (v/v) DMSO.xa0 Cell culture tested DMSO is available as ATCC Catalog No. 4-X.

Culture Conditions:
Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5%
Name of Depositor: DA Galloway
Deposited As: Mus musculus
References:

Halbert CL, et al. The E7 gene of human papillomavirus type 16 is sufficient for immortalization of human epithelial cells. J. Virol. 65: 473-478, 1991. PubMed: 1845902

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

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