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微信小章| 产品名称： | 7AC5/EYFP |
|---|---|
| 商品货号： | TS195046 |
| Organism： | Mus musculus, mouse |
| Tissue： | inner cell mass |
| Cell Type： | embryonic stem cell |
| Product Format： | frozen |
| Culture Properties： | adherent |
| Biosafety Level： | 2 xa0Cells contain CMV 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. |
| Age： | embryo |
| Gender： | male |
| Strain： | 129X1 x 129S1 |
| Applications： | SCRC-1033 has been shown to be germline competent, stains positive for pluripotency markers and alkaline phosphatase activity, and survives puromycin selection at concentrations of up to 0.004 mg/mL. |
| Storage Conditions： | liquid nitrogen vapor phase |
| Clinical Data： | male |
| Comments： | 7AC5/EYFP is a yellow fluorescent variant of R1 ES cells (ATCC SCRC-1011). The fluorescent variant was generated by the random integration of EYFP into R1 ES cells using co-electroporation with a circular selectable marker containing vector pPGK Puro. The vector is driven by a CMV immediate early enhancer coupled to the chicken beta-actin promoter and first intron. SCRC-1033 has been shown to be germline competent, stains positive for pluripotency markers and alkaline phosphatase activity, and survives puromycin selection at concentrations of up to 0.004 mg/mL. |
| Complete Growth Medium： | Grow ES cells in Mouse ES Cell Basal Medium (ATCC SCRR-2011) that has been supplemented with the following components: 1. 0.1 mM 2-mercaptoethanol (Life Technologies Cat. No. 21985-023) 2. 1,000 U/mL mouse leukemia inhibitory factor (LIF) (Millipore Cat. No. ESG1107) 3. 10% to 15% ES-Cell Qualified FBS (ATCC® SCRR-30-2020) or an ES cell qualified serum replacement Complete Growth Medium for Mouse ES Cells is stable for 14 days when stored at 2°C to 8°C. |
| Subculturing： | Subculturing Procedure Note: To insure the highest level of viability, pre-warm media and Trypsin/EDTA to 37ºC before adding to cells. Volumes used in this protocol are for T75 flasks. Proportionally adjust the volumes for culture vessels of other sizes. A split ratio of 1:4 to 1:7 is recommended. Feeder Cell Preparation for Subcultures
Dissociation and Transfer of ES Cells
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| Cryopreservation： | Freeze medium: Complete growth medium supplemented with an additional 10% FBS and 10% DMSO Storage temperature: liquid nitrogen vapor phase |
| Culture Conditions： | Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C |
| Name of Depositor： | A Nagy |
| Deposited As： | Mus musculus |
| References： | Matise M, et alProduction of targeted embryonic stem cell clonesIn: Matise M, et alGene Targeting: A Practical ApproachOxfordOxford University Press101-132, 1999 Hadjantonakis AK, et al. Generating green fluorescent mice by germline transmission of green fluorescent ES cells. Mech. Dev. 76: 79-90, 1998. PubMed: 9867352 Hadjantonakis AK, Nagy A. FACS for the isolation of individual cells from transgenic mice harboring a fluorescent protein reporter. Genesis 27: 95-98, 2000. PubMed: 10951501 Hadjantonakis AK, Nagy A. The color of mice: in the light of GFP-variant reporters. Histochem. Cell Biol. 115: 49-58, 2001. PubMed: 11219608 Hadjantonakis AK, et al. Embryonic stem cells and mice expressing different GFP variants for multiple non-invasive reporter usage within a single animal. BMC Biotechnol. 2: 11, 2002. PubMed: 12079497 |