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Human Mesenchymal Stem Cell Multi-Color Flow Kit

R&D Systems, Inc. a Bio-Techne Brand

Key Product Details

Verification of Mesenchymal Stem/Stromal Cell Identity by Analysis of Mesenchymal Stem/Stromal Cell Marker Expression.
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FMC002
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Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, the starting population of cells is characterized for MSC markers using the following procedure:

  • Incubate the cells with the provided antibodies
  • Wash the cells
  • Resuspend in Flow Cytometry Staining Buffer
  • Analyze by flow cytometry
 
 

Reagents Provided

Reagents Supplied in the Human Mesenchymal Stromal Cell Multi-Color Flow Cytometry Kit (Catalog # FMC002):

  • Flow Cytometry Staining Buffer (100 mL)

Positive Markers

  • Mouse IgG1 Anti-Human CD105-PerCP (Clone 166707)
  • Mouse IgG1 Anti-Human CD146-CFS (Clone 128018)
  • Mouse IgG2A Anti-Human CD90-APC (Clone Thy-1A1)

Negative Markers

  • Mouse IgG1 Anti-Human CD45-PE (Clone 2D1)

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

This protocol has been tested using bone marrow- and/or adipose tissue-derived MSCs. If using a different tissue source or cell line, the protocol below may need to be optimized.

 

 

 

 

 

 
  1. Wash the cells with Flow Cytometry Staining Buffer.
    Perform a cell count.
  2. 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.
  3. Transfer approximately 100 µL of the Fc receptor-blocked cells (about 1 x 106 cells) into a 5 mL flow cytometry tube.
  4. Add 10 µL of each fluorochrome-conjugated antibody or each corresponding isotype control antibody.
  5. Incubate the mixture for 30-45 minutes at room temperature in the dark.
  6. Centrifuge the samples at 300 x g for 5 minutes.
  7. Wash the cells with 2 mL Flow Cytometry Staining Buffer.
  8. Resuspend the cell pellet in 200-400 µL of Flow Cytometry Staining Buffer.
  9. Analyze expression of MSC markers simultaneously by flow cytometry.

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    Name: Anonymous
    Verified Customer | Posted 09/11/2018
    Human Mesenchymal Stem Cell Multi-Color Flow Kit FMC002

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FAQs for Human Mesenchymal Stem Cell Multi-Color Flow Kit

  • Will  the Human Mesenchymal Stem Cell Multi-Color Flow Kit (catalog # FMC002) or Mouse Mesenchymal Stem Cell Multi-Color Flow Kit (catalog # FMC003) work for identificaiton of Rat Menchymal stem cells?

    The Human Mesenchymal Stem Cell Multi-Color Flow Kit (catalog # FMC002) or Mouse Mesenchymal Stem Cell Multi-Color Flow Kit (catalog # FMC003) will not work for identificaiton of Rat Mesenchymal stem cells.

Product Documents for Human Mesenchymal Stem Cell Multi-Color Flow Kit

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.