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 - Complete: all you need to isolate and quantify rare EPC populations
- Time-saving: pre-titrated antibody cocktail minimizes assay setup time
- Ultimate convenience: combine with the Express Mode for automated flow cytometric analysis on the MACSQuant® Analyzer
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| Overview |
The EPC Enrichment and Enumeration Kit is designed for the enumeration of circulating EPCs (cEPCs) from peripheral blood, cord blood, or leukapheresis products as well as EPCs from bone marrow. To ensure reliable enumeration results, the EPCs are enriched during the procedure to increase the sensitivity of the flow cytometric analysis.
The kit contains all reagents for:
- red blood cell lysis,
- EPC enrichment,
- blocking of Fc receptor–mediated non-specific labeling of non-target cells,
- enumeration of EPCs based on expression of CD34, CD133, and CD309,
- exclusion of CD14+ cells,
- exclusion of dead cells,
- isotype controls.
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| Details |
Background information
Endothelial progenitor cells (EPCs) are defined by the expression of CD34, CD133, and CD309 (VEGFR-2/KDR)1,2 and have been enumerated based on the expression of these markers by several studies. They are considered a measurable parameter for the assessment of risk factors for various diseases3–5 as well as for the capacity for vascular repairing. cEPC numbers were correlated with, for example,
- certain phases during congestive heart failure6,
- acute myocardial infarction7,
- risk factors for atherosclerosis5,
- risk factors for cardiovascular disease8,
- physical training9,
- cessation of smoking10.
The EPC Enrichment and Eunmeration Kit contains everything needed for EPC enrichment followed by enumeration with CD34, CD133/2, CD309, CD14 fluorochrome-conjugated monoclonal antibodies. |
| Columns |
| MS Columns or MACSQuant Columns |
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| Figure 1 |
| Identification of CD34+ cells and CD133 specificity. Leukocytes after discrimination of debris and platelets and exclusion of dead cells and monocytes are shown. Cells were stained with CD34-FITC and Mouse IgG2b-PE representing the isotype control for CD133 staining. Cells were analyzed by flow cytometry. |
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| Figure 2 |
| Identification of CD34+CD133+ cells in the pre-enriched EPC sample. Leukocytes after discrimination of cell debris and platelets and exclusion of dead cells and monocytes are shown. Cells were stained with CD34-FITC and CD133/2 (293C3)-PE and analyzed by flow cytometry. The population in R4 was used for further analysis of CD133 and CD309 expression shown in figures 3 and 4. |
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| Figure 3 |
| Enumeration of CD34+CD133+CD309 (VEGFR-2/KDR)+ EPCs in the pre-enriched EPC sample. Leukocytes after discrimination of debris and platelets, exclusion of dead cells and monocytes are shown. Additionally, data were gated on CD34+ cells. Cells were stained with CD309 (VEGFR-2/KDR)-APC and CD133/2 (293C3)-PE and analyzed by flow cytometry. |
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| Figure 4 |
| Isotype control of CD309 staining in the pre-enriched EPC sample. Leukocytes after discrimination of debris and platelets, exclusion of dead cells and monocytes are shown. Additionally, data were gated on CD34+ cells. Cells were stained with CD133/2 (293C3)-PE and Mouse IgG1-APC representing the isotype control for CD309 (VEGFR-2/KDR) staining. Cells were analyzed by flow cytometry. |
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| Products |
| EPC Enrichment and Enumeration Kit, human |
- for 20 tests (1)
Download datasheet
130-093-477
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| (1) One test corresponds to 20.5 mL whole blood, cord blood or bone marrow or to 2×108 white blood cells for leukapheresis product. |
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| References |
| 1. Peichev, M. et al. (2000) Blood 95: 952–958. |
| 2. Rafii, S. and Lyden, D. (2003) Nat. Med. 9: 702–712. |
| 3. Hill, J. et al. (2003) N. Engl. J. Med. 348: 593–600. |
| 4. Urbich, C. and Dimmeler, S. (2005) Kidney Int. 67: 1672–1676. |
| 5. Werner, N. and Nickenig, G. (2006) Arterioscler. Thromb. Vasc. Biol. 26: 257–266. |
| 6. Valgimigli, M. et al. (2004) Circulation 110: 1209–1212. |
| 7. Massa, M. et al. (2005) Blood 105: 199–206. |
| 8. Vasa, M. et al. (2001) Circ. Res. 89: e1–e7. |
| 9. Steiner, S. et al. (2005) Atherosclerosis 181: 305–310. |
| 10. Kondo, T. et al. (2004) Arterioscler. Thromb. Vasc. Biol. 24: 1442–1447. |
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