Mouse Hematopoietic Progenitor Cell Multi-Color Flow Kit
Mouse Hematopoietic Progenitor Cell Multi-Color Flow Kit Summary
Kit Summary
Why is it Important to Verify Hematopoietic Progenitor Cell Identity using Established Markers?
Researchers use different techniques for isolating, culturing, and differentiating mouse hematopoietic progenitor cells. See Details
Variations in experimental approaches as well as differences in the starting cell population may account for experimental variability and contradictory data that have been published in the stem cell field. One way to minimize experimental variability is to clearly define the starting cell population by using a functional or phenotypic assay.
Verification of hematopoietic progenitor cell identity by multi-color flow cytometry:
- Uses expression of established markers to verify progenitor cell identity and multipotency.
- Verifies a healthy, multipotent starting progenitor cell population to increase consistency between studies and reduce unwanted experimental variability.
- Does not require the use of secondary antibodies.
Kit Contents
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This kit contains the following three fluorochrome-conjugated primary antibodies and staining buffer for flow cytometry: See Details
- PE-conjugated Rat Anti-Mouse CD48/SLAMF2 Monoclonal Antibody (Clone 331504; rat IgG1)
- APC-conjugated Rat Anti-Mouse SLAM/CD150 Monoclonal Antibody (Clone 459911; rat IgG2A)
- CFS-conjugated Goat-Anti Mouse 2B4/CD244/SLAMF4 Affinity-purified Polyclonal Antibody; goat IgG)
- Staining Buffer (100 mL)
The quantity of each antibody in this kit is sufficient to process 25 flow cytometry samples at the recommended concentrations.
Data Examples
 View Larger Image | Verification of Mouse Bone Marrow-derived Hematopoietic Progenitor Cell Identity using Multi-Color Flow Cytometry. Mouse bone marrow-derived cells were stained using reagents supplied in the Mouse Hematopoietic Progenitor Cell 3-Color Flow Kit (Catalog # FMC005). Cells were simultaneously analyzed for the expression of multipotency markers including CD48/SLAMF2, SLAM/CD150, and 2B4/CD244/SLAMF4 by flow cytometry. (A) The dot plot shows the relative CD48/SLAMF2+ and 2B4/CD244/SLAMF4- population from (B) SLAM/CD150+-gated bone marrow-derived cells. Quadrants were set based on isotype controls. |
 View Larger Image | Verification of Mouse Splenocyte Identity using Multi-Color Flow Cytometry. Mouse splenocytes were stained using reagents supplied in the Mouse Hematopoietic Progenitor Cell 3-Color Flow Kit (Catalog # FMC005). The cells were stained with the indicated antibodies against multipotency markers (filled histograms) or the corresponding isotype control antibodies (open histograms). |
Specifications
Product Datasheets
Assay Procedure
Refer to the product datasheet for complete product details.
Reagents Provided
Reagents Supplied in the Mouse Hematopoietic Progenitor Cell 3-Color Flow Kit (Catalog # FMC005)
- PE-conjugated Rat Anti-Mouse CD48 Monoclonal Antibody (Clone 331504; rat IgG1)
- APC-conjugated Rat Anti-Mouse CD150 Monoclonal Antibody (Clone 459911; rat IgG2A)
- CFS-conjugated Goat-Anti Mouse CD244 Affinity-purified Polyclonal Antibody; goat IgG)
- Rat IgG1 Isotype Control
- Rat IgG2A Isotype Control
- Goat IgG Isotype Control
- 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
- Perform a cell count on cells that have been washed with Flow Cytometry Staining Buffer.
- 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 106 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 MSC markers simultaneously by flow cytometry.
FAQs
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