MilliTrace Nanog GFP Reporter Mouse Embryonic Stem Cell Kit

MilliTrace Nanog GFP Reporter Mouse Embryonic Stem Cell Kit

1 kit

MilliTraceTM Nanog GFP Reporter Mouse Embryonic Stem Cell Kit (Catalog No. SCR089) provides ready-to-use mouse embryonic stem cells that express the humanized mulleri green fluorescent protein (hmGFP) reporter gene under the control of the mouse Nanog promoter along with expansion medium to help maintain expression of the transgene.

MilliTrace Nanog GFP Reporter Mouse Embryonic Stem Cells were generated by transfection of C57/BL6 mouse embryonic stem cells with a proprietary bicistronic plasmid construct containing hmGFP under the control of the mouse Nanog promoter. FACS analyses of stable transfectants indicate that greater than 90% of the cells express GFP at high levels even after 10 passages when cultured at normal proliferation conditions. MilliTrace Nanog GFP Reporter Mouse Embryonic Stem Cells display the immunochemical staining properties of pluripotent stem cells; they are positive for alkaline phosphatase and Nanog-hmGFP expression is co-localized with Oct-4, Sox-2, and SSEA-1 expression. Upon differentiation, hmGFP is down-regulated and cells have the capacity to differentiate into multiple lineages, including cardiomyocytes and neurons. Under neuronal differentiation conditions, cells can be differentiated to βIII-tubulin positive neurons (GFP+βIII-tubulin+) (Use Mouse Embryonic Stem Cell Neurogenesis Kit, Catalog No. SCR101).

Nanog is an important marker of undifferentiated embryonic stem cells and, along with Oct-4 and Sox-2, is an important regulator of stem cell pluripotency. Studies have shown that down-regulation of the Nanog promoter coincides with the loss of pluripotency and progression towards a differentiated state. Nanog GFP Reporter Mouse Embryonic Stem Cell Kit provides a quick non-invasive method by which to monitor the expression of Nanog in pluripotent embryonic stem cells. The cells can be used to facilitate studies elucidating the role of Nanog and other factors in the maintenance and self-renewal of embryonic stem cells. This kit can also be used to identify factors involved in embryonic stem cell differentiation.

For Research Use Only. EMD Millipore does not recommend that a user derive clones from cells provided with this kit. However, should a user decide to do so, they may need a license to one or more third party patents and it is incumbent upon that user to determine whether and if so which third party patents need to be licensed.

Nanog GFP reporter mouse embryonic stem cells have been validated for expression of hmGFP in the pluripotent state, for high level of expression of alkaline phosphatase, Oct-4, Sox-2, and SSEA-1, and for their self-renewal and multi-lineage differentiation capacities. Approximately 80% cells display normal karyotype as assessed by G-banding of twenty metaphase cells. Cells tested negative for mycoplasma.

Mouse Embryonic Stem Cells

• Cells/Isolation
• Maintenance & Expansion

Embryonic Stem Cells

• MilliTrace Nanog GFP Reporter Mouse Embryonic Stem Cells, 1 x 10⁶ viable cells
• MilliTrace Mouse Embryonic Stem Cell Expansion Medium (Catalog No. SCM042)

· MilliTrace Nanog GFP Reporter Mouse Embryonic Stem Cells (Part No. SCC089) should be stored in Liquid Nitrogen. We recommend that the cells be used within ten passages.
· EmbryoMax Complete ES Cell Medium with 15% FBS and LIF (Part No. ES-101-B) should be stored at -20°C until expiration date on the label. Upon thawing the medium should be stored at 2-8°C and given a 1-month expiration dating.
· 5 mg/mL Puromycin Solution (Part No. CS201361) should be stored in working aliquots at -20°C for up to 2 years.

MilliTrace Nanog GFP Reporter Mouse Embryonic Stem Cell Kit