HR1 Cell Line

HR1 cells are HEK 293 cells stably expressing conditional kinase <U+2206>Raf-1:ER* from the pCMV Neo Myc plasmid. <U+2206>Raf-1:ER* (also known as <U+2206>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.

Contributors

Inventor Institute
Simon Cook Babraham Institute
Cat. #: 153329
Tool sub type: Continuous
Unit size: 1×10^6 cells / vial
Research Fields: Apoptosis and autophagy;Cancer;Cell biology;Cell signaling and signal transduction;Immunology
Organism: Human
Tissue: Kidney
Growth properties: Adherent cell line
Alternate name: RAF proto-oncogene serine/threonine-protein kinase (EC:2.7.11.1), Proto-oncogene c-RAF, cRaf, Raf-1
Product 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 β-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.
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.
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
Parental cell line: HEK 293
Disease: Cancer
Format: Frozen
Shipping conditions: Dry ice
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).
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(1…

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