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Contributor Information

  • Name Simon Cook
  • Institute Babraham Institute

Tool Details

  • Tool name: HR1 Cell Line
  • Alternate names: RAF proto-oncogene serine/threonine-protein kinase (EC:2.7.11.1), Proto-oncogene c-RAF, cRaf, Raf-1
  • Tool type: Cell Lines
  • Tool sub-type: Continuous
  • Parental cell line: HEK 293
  • Organism: Human
  • Tissue: Kidney
  • Disease: Cancer; Inflammation
  • Growth properties: Adherent cell line
  • Conditional: Yes
  • Conditional description: Stable transfectant, conditional functionality: c-Raf1 kinase activity and ERK1/2 signalling pathway activated following 4-hydroxytamoxifen (4-HT) treatment of cells.
  • Description: HR1 cells are HEK 293 cells stably expressing conditional kinase Raf-1:ER* from the pCMV Neo Myc plasmid. Raf-1:ER* (also known as CRAF:ER*) consists of the isolated kinase domain of c-Raf1 fused in-frame to a modified form of the hormone binding domain of the estrogen receptor (hbER*) that can be de-repressed by 4-hydroxytamoxifen (4-HT) but not ?-estradiol. In this case the * refers to a point mutation that ablates estradiol binding but allows 4-HT binding. Activation of Raf-1:ER* leads to the selective activation of the ERK1/2 pathway. This cell line can be used to study the cellular role and factors impacting on the ERK1/2 signalling pathway such as gene expression, cell proliferation, cell cycle arrest and cell death. The cells are G418 resistant. Conditional kinase activation of Raf-1:ER* can be induced with 100nM 4-HT.
  • Research area: Cancer; Apoptosis and autophagy; Cell biology; Cell signaling and signal transduction; Immunology
  • Production details: HEK 293 cells were transfected with the pCMV Neo Myc plasmid expressing â?‚ˆ?‚†Raf-1:ER*. Stable transfectants were selected by neomycin resistance using G418 (Geneticin) and ring cloning.
  • Cellosaurus ID: CVCL_2676

  • For Research Use Only

Target Details

  • Target: c-Raf1

Application Details

Handling

  • Format: Frozen
  • Growth medium: Phenol red-free Dulbecco's modified Eagle's medium (DMEM) high glucose version, 2 mM L-glutamine, 10% fetal bovine serum (FBS), 400 ?g/ml G418 (Geneticin).
  • Shipping conditions: Dry ice

Documentation

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References

  • •  Gilley et al. 2009. Cell Signal. 21(6):969-77. PMID: 19249353.
  • •  ERK1/2, but not ERK5, is necessary and sufficient for phosphorylation and activation of c-Fos.
  • •  Wiggins et al. 2007. Cell Signal. 19(12):2605-11. PMID: 17884340.
  • •  c-Cbl is not required for ERK1/2-dependent degradation of BimEL.
  • •  Ewings et al. 2007. EMBO J. 26(12):2856-67. PMID: 17525735.
  • •  ERK1/2-dependent phosphorylation of BimEL promotes its rapid dissociation from Mcl-1 and Bcl-xL.
  • •  Boughan et al. 2006. J Biol Chem. 281(17):11637-48. PMID: 16513653.
  • •  Nucleotide-binding oligomerization domain-1 and epidermal growth factor receptor: critical regulators of beta-defensins during Helicobacter pylori infection.