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| Stemgent® Mouse ES/iPS Cell Characterization Set |
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| Description |
The Stemgent™ Mouse ES/iPS Cell Characterization Set is a specific and sensitive, stem cell-verified tool designed to analyze the undifferentiated and differentiated status of mouse embryonic stem (ES) and induced pluripotent stem (iPS) cells. The set includes the Stemgent™ AP Staining Kit to measure alkaline phosphatase (AP) activity, and three ES cell-verified Stemgent™ antibodies to detect the mouse ES/iPS cell surface stage-specific embryonic antigen-1 (SSEA-1), and the intracellular transcription factors, Nanog and Oct4. The Stemgent™ Mouse ES/iPS Cell Characterization Set is quality tested by monitoring the AP staining activity of mouse ES cells, and assessing the undifferentiated/differentiated cell states through the expression of SSEA-1, Oct4, and Nanog by immunocytochemistry (ICC) and/or flow cytometry (FC).
Mouse ES and iPS cells are of extensive research interest because of their capacity for unlimited proliferation in vitro and their unique differentiation potential for future therapeutic use1,2. While the undifferentiated/pluripotent state of ES cells can best be defined functionally, the identification and characterization of ES and iPS cells by their unique biomarker(s) are extremely important to assess the quality of the cells in culture and the efficiency of cell reprogramming. The undifferentiated state of ES cells and iPS cells can be characterized by a high level of AP expression along with the expression of the mouse pluripotency markers SSEA-1, Nanog, and Oct4.
Alkaline Phosphatase (AP) is a hydrolase enzyme responsible for dephosphorylating molecules such as nucleotides, proteins, and alkaloids under alkaline conditions. When fixed ES cells or iPS cells are stained with AP, undifferentiated cells appear red or purple, whereas differentiated cells appear colorless, making this a simple, sensitive, and rapid assay for the verification of the pluripotent state3,4.
SSEA-1, a carbohydrate antigen, is expressed on the surface of preimplantation-stage mouse embryos at the eight-cell stage5. It is strongly expressed by undifferentiated mouse ES cells6. Upon differentiation, mouse ES cells are characterized by the loss of SSEA-1 expression7.
Both Nanog and Oct4 are transcription factors that have been associated with an undifferentiated phenotype in embryonic ES/iPS cell8-10. A critical amount of Nanog and Oct4 expression is required to sustain stem-cell pluripotency. These key factors form a regulatory network to support or limit each other's expression level and maintain the properties of ES cells. When ES cells are induced to differentiate, Nanog and Oct4 are downregulated, which has proven to be essential for a proper and divergent developmental program11,12. Functionally, Nanog and Oct4 work together with other key reprogramming factors (Sox2, Lin28, Klf4 and C-Myc) to reprogram mouse and human fibroblasts and generate induced pluripotent stem (iPS) cells13-15. |
| Specifications and technical documents |
| Specification SheetsMaterial Safety Data SheetsProtocols
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| Figure 1 |
| SSEA-1: Flow Cytometry Analysis of Mouse ES Cell Line R1. |
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| Figure 2 |
| Oct4: Immunocytochemistry Analysis of Mouse ES Cell Line R1. |
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| Figure 3 |
| AP staining of Mouse ES cell line R1. |
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| Details |
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| Products |
| Stemgent Mouse ES/iPS Cell Characterization Set |
- 50 assays
- Stemgent Catalog #00-0016
130-095-612
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| References |
| 1. Evans et al. (1981) Nature 292: 154–156. |
| 2. Martin et al. (1981) Proc Natl Acad Sci USA 78: 7634–7638. |
| 3. Saito et al. (2004) Hum Cell. 17 (3): 107–115. |
| 4. Mitalipov et al. (1994) Ontogenez. 25 (6): 19–27. |
| 5. Solter and Knowles (1978) Proc. Natl. Acad. Sci. USA 75: 5565–5569. |
| 6. Matsui et al. (1992) Cell 70: 841–847. |
| 7. Solter and Knowles (1979) Curr Top Dev Biol. 13: 139–165. |
| 8. Nichols et al. (1998) Cell 95: 379–391. |
| 9. Mitsui et al. (2003) Cell 113: 631–642. |
| 10. Chambers et al. (2003) Cell 113 (5): 643–655. |
| 11. Pan and Thomson (2007) Cell Res. 17: 42–49. |
| 12. Kim et al. (2008) Cell 132: 1049–1061. |
| 13. Takahashi and Yamanaka (2006) Cell 126: 663–676. |
| 14. Takahashi et al. (2007) Cell 131: 861–872. |
| 15. Yu et al. (2007) Science 318: 1917–1920. |
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