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微信小章| 产品名称： | PERK-KO-DR |
|---|---|
| 商品货号： | TS176200 |
| Organism： | Mus musculus, mouse |
| Tissue： | embryo; fibroblast |
| Cell Type： | fibroblast |
| Product Format： | frozen |
| Morphology： | fibroblast-like |
| Culture Properties： | adherent |
| Biosafety Level： | 2
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. |
| Age： | embryo, 13.5 days gestation |
| Applications： | ER stress; unfolded protein response signaling pathway |
| Storage Conditions： | liquid nitrogen vapor phase |
| Images： |  |
| Derivation： | The mouse embryonic fibroblast (MEF) cell line, PERK-KO-DR (CRL-2976), was established from a 13.5 day-old PERK -/- mouse embryo by SV-40 immortalization. These cells lack the Endoplasmic Reticulum (ER) localized stress induced kinase, PERK, which mediates signaling from the stressed ER to the translational apparatus by phosphorylating eukaryotic initiation factor 2a (eIF2a). Phosphorylation of eIF2a results in a global inhibition of protein synthesis. PERK is a major regulator of the unfolded protein response (UPR), and as such, these PERK-KO-DR cells are very useful to investigate ER stress and UPR signaling in a variety of contexts. DR-Wildtype cells, ATCC CRL-2977, are available for use as a control. In addition, GCN2 knockout MEF cell line ATCC CRL-2978 and CHOP knockout MEF cell line ATCC CRL-2979 are two related important tools for ER stress and UPR studies. |
| Genes Expressed： | protein kinase RNA-like endoplasmic reticulum kinase (PERK), not expressed |
| Cellular Products： | protein kinase RNA-like endoplasmic reticulum kinase (PERK), not expressed Ref   Harding HP, et al. Perk is essential for translational regulation and cell survival during the unfolded protein response. Mol. Cell 5(5): 897-904, 2000. PubMed: 10882126 |
| Comments： | The mouse embryonic fibroblast (MEF) cell line, PERK-KO-DR (CRL-2976), was established from a 13.5 day-old PERK -/- mouse embryo by SV-40 immortalization. These cells lack the Endoplasmic Reticulum (ER) localized stress induced kinase, PERK, which mediates signaling from the stressed ER to the translational apparatus by phosphorylating eukaryotic initiation factor 2a (eIF2a). Phosphorylation of eIF2a results in a global inhibition of protein synthesis. PERK is a major regulator of the unfolded protein response (UPR), and as such, these PERK-KO-DR cells are very useful to investigate ER stress and UPR signaling in a variety of contexts. DR-Wildtype cells, ATCC CRL-2977, are available for use as a control. In addition, GCN2 knockout MEF cell line ATCC CRL-2978 and CHOP knockout MEF cell line ATCC CRL-2979 are two related important tools for ER stress and UPR studies. |
| 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:
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| Subculturing： | Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
Subcultivation ratio: A subcultivation ratio of 1:10 to 1:30 twice weekly is recommended.
Medium renewal: every 2 to 3 days
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney,xa05th edition, published by Wiley-Liss, N.Y., 2005. |
| Cryopreservation： | Fetal bovine serum (FBS), 90%; DMSO, 10%. 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： | D Ron and H Harding |
| Year of Origin： | 2000 |
| References： | Harding HP, et al. Protein translation and folding are coupled by an endoplasmic-reticulum-resident kinase. Nature 397(6716): 271-274, 1999. PubMed 9930704 Harding HP, et al. Perk is essential for translational regulation and cell survival during the unfolded protein response. Mol. Cell 5(5): 897-904, 2000. PubMed: 10882126 Harding HP, et al. Diabetes mellitus and exocrine pancreatic dysfunction in perk -/- mice reveals a role for translational control in secretory cell survival. Mol. Cell 7(6): 1153-1163, 2001. PubMed: 11430819 Kimball SR, et al. Mammalian stress granules represent sites of accumulation of stalled translation initiation complexes. Am. J. Physiol. Cell Physiol. 284(2): C273-C284, 2003. PubMed: 12388085 Hay RJ, Caputo JL, Macy, ML, Eds. (1992) ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC. Caputo JL. 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. |