Human Hematopoietic Progenitor Cell Multi-Color Flow Kit
R&D Systems, part of Bio-Techne | Catalog # FMC019
Key Product Details
Summary for Human Hematopoietic Progenitor Cell Multi-Color Flow Kit
Kit Summary
| HSC Differentiation &Lineage Marker Schematic View Interactive Pathway |
The definitive hematopoietic system is made up of all adult blood cell types including megakaryocytes, erythrocytes, and cells of the myeloid and lymphoid lineages. All of these cells are derived from multipotent hematopoietic stem cells (HSCs) through a succession of precursors with progressively limited potential. Hematopoietic stem cells are tissue-specific stem cells that exhibit remarkable self-renewal capacity and are responsible for the life-long maintenance of the hematopoietic system. HSCs are rare cells that reside in adult bone marrow where hematopoiesis is continuously taking place. They can also be found in cord blood, fetal liver, adult spleen, and peripheral blood. R&D Systems offers several products for studying hematopoietic lineage cells including serum-free media, lineage depletion antibodies and kits, and reagents for performing colony forming cell (CFC) assays.
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Identification of Human Hematopoietic Progenitor Cells using Multi-Color Flow Cytometry. Human umbilical cord blood cells were stained using reagents supplied in this kit. Cells were simultaneously analyzed for the expression of multipotency markers. Cells negative for CD11b and negative/low for CD38 (boxed area in A) were gated and assessed for positive expression of CD34 and SCF R/CD117 (upper right quadrant in B). Quadrants were set based on isotype controls. |
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Identification of Human Hematopoietic Progenitor Cells using Multi-Color Flow Cytometry. Human umbilical cord blood cells were stained with the indicated antibodies included in this kit. Representative expression of each analyte in the total umbilical cord blood cell population (filled histogram) over the isotype control (open histogram). |
Assay Procedure
Refer to the product datasheet for complete product details.
Reagents Provided
Reagents Supplied in the Human Hematopoietic Progenitor Cell Multi-Color Flow Kit (Catalog # FMC019)
- APC-conjugated Mouse Anti-Human CD34 (Clone QBEnd10; Mouse IgG1)
- PE-conjugated Mouse Anti-Human SCF R/CD117 (Clone 47233; Mouse IgG1)
- PerCP-conjugated Mouse Anti-Human CD38 (Clone 240742; Mouse IgG2A)
- CFS-conjugated Mouse Anti-Human CD11b (Clone 238446,IgG2B)
- APC-conjugated Mouse IgG1 Isotype Control (Clone 11711)
- PE-conjugated Mouse IgG1 Isotype Control (Clone 11711)
- PerCP-conjugated Mouse IgG2A Isotype Control (Clone 20102)
- CFS-conjugated Mouse IgG2B Isotype Control (Clone 133303)
- Flow Cytometry Staining buffer
Other Supplies Required
Reagents
- Fc Receptor Blocking Reagents
Materials
- Flow Cytometry/FACS™ Tubes (5 mL round-bottom polystyrene tubes)
- Pipette Tips and Pipettes
Equipment
- Centrifuge
- Vortex
Procedure Overview
- Resuspend the cells in Flow Cytometry Staining Buffer at 1 x 105 cells/100 µL.
- Add Fc receptor blocking reagents. If using excess pre-immune IgG to block Fc receptor, the excess IgG does not need to be washed from the cells following the incubation period.
- Transfer approximately 100 µL of the Fc receptor-blocked cells (about 1 x 105 cells) into a 5 mL flow cytometry tube.
- Add 10 µL of each antibody or each corresponding isotype control antibody.
- Incubate the mixture for 30-45 minutes at room temperature in the dark.
- Centrifuge the samples at 300 x g for 5 minutes.
- Wash the cells with 2 mL of Flow Cytometry Staining Buffer.
- Resuspend the cell pellet in 200-400 µL of Flow Cytometry Staining Buffer.
- Analyze the expression of hematopoietic progenitor cell markers simultaneously by flow cytometry.
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