Mesechymal Cell Kit

Details for Product No. ABIN2669909, Supplier: Log in to see
Reactivity
Human
Application
Flow Cytometry (FACS)
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Purpose Reagent kit for identification of human multipotent mesenchymal stromal cells by flow cytometry.
Sample Type Mesenchymal Cells
Detection Method Fluorometric
Components The kit is composed by a combination of differents murine monoclonal antibodies into 3 vials, A, B and C.
  • Vial A: CD90 FITC, CD105 PE, CD45 PerCP, CD19 PerCP, HLA-DR PerCP, CD14 PerCP, CD34 PerCP, CD29 APC, 50 Tests
  • Vial B: CD44 FITC, CD166 PE, CD45 PerCP, CD19 PerCP, HLA-DR PerCP, CD14 PerCP, CD34 PerCP, CD73 APC, 50 Tests
  • Vial C: Isotype control IgG1 FITC, Isotype control IgG1 PE, Isotype control IgG1 PerCP, Isotype control IgG1, 50 Tests
In addition the kit includes four aliquots of antibody to perform the compensation: CD90 FITC, CD166 PE, CD44 PerCP and CD29 APC.
Material not included
  • Plastic test tubes (12 x 75 mm)
  • Calibrated standard pipets (20 μL, 100 μL, 2 mL) and tips
  • Vortex mixer
  • Timer
  • Flow cytometer
  • analysis software
Application Notes Optimal working dilution should be determined by the investigator.
Comment

It is recommended for identification and phenotyping of cultured human multipotent mesenchymal stromal cells by flow cytometry.Samples were tested on Flow Cytometry Samples can be run up to 3 hours after lysis.

Sample Volume 5 μL
Assay Time < 1 h
Protocol Cell surface Protocol:
1. Transfer up 1x106 cells to a 12 x 75 mm polystyrene test each of the three tubes A, B and C.
2. Centrifuge at 540g for 5 minutes. Gently aspirate the supernatant and discard it leaving approximately 50 μL of fluid. Add 50 μL of buffer and mix gently with a vortex mixer.
3. Add 20 μL of vial A into tube A, 20 μL of vial B into tube B and 20 μL of vial C into tube C. Mix gently with a vortex mixer. The 20 μL is a guideline only, the optimal volume should be determined by the individual laboratory.
4. Incubate in the dark at room temperature (20-25 °C) for 15 minutes or at 4 °C for 30 minutes.
5. Add 2 mLof buffer and mix gently with a vortex mixer
6. Centrifuge at 540g for 5 minutes. Gently aspirate the supernatant without disturbing the cell pellet and discard it leaving approximately 50 μL of fluid.
7. Resuspend pellet adding 50 μL of buffer.
Reagent Preparation
  • Reagent requeriments
  • Prepare a solution containing PBS pH 7,2, 0,5 % BSA and 2 mM EDTA, Ca2+ and Mg2+ free. Store at 2-8° C
Restrictions For Research Use only
Preservative Sodium azide
Precaution of Use Reagents contain sodium azide. Sodium azide under acid conditions yields hydrazoic acid, an extremely toxic compound. Azide compounds should be diluted with running water before being discarded. These conditions are recommended to avoid deposits in plumbing where explosive conditions may develop.
Do not pipet by mouth.
Samples should be handled as if capable of transmitting infection. Appropriate disposal methods should be used.
The sample preparation procedure employs a fixative (formaldehyde). Contact is to be avoided with skin or mucous membranes.
The flow cytometer should be compensated specifically for this kit as indicated in Annex
7. Do not use antibodies beyond the stated expiration dates of the products
Cultured MSCs have to be dissociated with Trypsin and used immediately
Handling Advice Light exposure should be avoided. Use dim light during handling, incubation with cells and prior to analysis.
Storage 4 °C
Background publications Dominici, Le Blanc, Mueller, Slaper-Cortenbach, Marini, Krause, Deans, Keating, Prockop, Horwitz: "Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement." in: Cytotherapy, Vol. 8, Issue 4, pp. 315-7, 2006 (PubMed).