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Human Mesenchymal Stem Cell Marker Antibody Panel

R&D Systems, part of Bio-Techne | Catalog # SC017

Contains 25 ug each of antibodies to Stro-1, CD90, CD106, CD105, CD146, CD166, CD44, CD19, and CD45.
R&D Systems, part of Bio-Techne

Key Product Details

Verification of Human Mesenchymal Stem/Stromal Cell Identity by Analysis of MSC Marker Expression.

Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, human mesenchymal stem cell marker expression can be assessed using the following procedure:

  • Incubate cells with the provided antibodies for 30 minutes
  • Wash the cells
  • Incubate the cells with fluorochrome-conjugated secondary antibodies
  • Analyze samples by flow cytometry
 

 

Reagents Provided

Reagents Supplied in the Human Multipotent Mesenchymal Stromal Cell Marker Antibody Panel (Catalog # SC017):

Positive MSC Markers

  • Mouse Anti-Human Stro-1 IgM gamma Monoclonal Antibody
  • Mouse Anti-Human CD44 IgG2A Monoclonal Antibody
  • Mouse Anti-Human CD-90 IgG2A Monoclonal Antibody
  • Mouse Anti-Human CD105 IgG1 Monoclonal Antibody
  • Mouse Anti-Human CD106 IgG1 Monoclonal Antibody
  • Mouse Anti-Human CD146 IgG1 Monoclonal Antibody
  • Mouse Anti-Human CD166 IgG1 Monoclonal Antibody

Negative MSC Markers

  • Mouse Anti-Human CD19 IGg1 Monoclonal Antibody
  • Mouse Anti-Human CD45 IGg1 Monoclonal Antibody

 

Other Supplies Required

Reagents

Materials

  • Flow Cytometry Tubes
  • Serological pipettes

Equipment

  • Benchtop centrifuge
 

 

Procedure Overview

Staining of Mesenchymal Stem Cell Surface Markers

Perform a cell count on harvested cells.

Resuspend cells in Flow Cytometry Staining Buffer.

Generate embryoid bodies (EB) from pluripotent stem cells

Aliquot 90 μl of the cells into 5 mL flow cytometry tubes.

Replace the KO-ES Medium with 10 mL of ITS/Fibronectin Media

Add 10 μL of antibody or isotype control (or a previously titrated amount).

Vortex and incubate for 30 minutes at room temperature.

Wash the attached cells twice with sterile PBS

Centrifuge samples at 300 x g for 5 minutes.

Wash the samples three times with Flow Cytometry Staining Buffer.

Resuspend each sample in 200 μL of Flow Cytometry Staining Buffer.

Transfer the cells to a 15 mL tube

Add 10 μL of a fluorochrome-conjugated secondary antibody (or a previously titrated amount).

Incubate for 30 minutes at room temperature in the dark.

Centrifuge the suspension for 5 minutes at 220 x g

Centrifuge the samples at 300 x g for 5 minutes.

Wash the samples with Flow Cytometry Staining Buffer.

Resuspend the cells in 200-400 μL of Flow Cytometry Staining Buffer.

Perform a cell count

Analyze the cells by flow cytometry.

Plate the cells at 1 x 105 cells/well in 500 µL of N-2 MAX/FGF Media onto Poly-L-ornithine/Fibronectin coated plates
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Product Documents for Human Mesenchymal Stem Cell Marker Antibody Panel

Certificate of Analysis

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