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Human Hematopoietic Progenitor Cell Multi-Color Flow Kit

R&D Systems, part of Bio-Techne | Catalog # FMC019

Contains conjugated antibodies to CD34-APC (Clone QBEnd10), CD38-PerCP (Clone 240742), CD11b-Fluorescein (Clone 238446), SCF R-PE (Clone 74233)
R&D Systems, part of Bio-Techne

Key Product Details

Identification of Human Hematopoietic Progenitor Cells using Multi-Color Flow Cytometry.
(2)

Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, hematopoietic progenitor cells are identified by flow cytometry using the following procedure:

  • Suspend cells in Flow Cytometry Staining Buffer
  • Incubate the cells with fluorochrome-conjugated antibodies or isotype controls
  • Wash the cells
  • Resuspend in Flow Cytometry Staining Buffer
  • Analyze by flow cytometry
 
 

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

 
  1. Resuspend the cells in Flow Cytometry Staining Buffer at 1 x 105 cells/100 µL.

 
  1. 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.

  1. Transfer approximately 100 µL of the Fc receptor-blocked cells (about 1 x 105 cells) into a 5 mL flow cytometry tube.

  1. Add 10 µL of each antibody or each corresponding isotype control antibody.
  2. Incubate the mixture for 30-45 minutes at room temperature in the dark.
 

  1. Centrifuge the samples at 300 x g for 5 minutes.

 
  1. Wash the cells with 2 mL of Flow Cytometry Staining Buffer.

  1. Resuspend the cell pellet in 200-400 µL of Flow Cytometry Staining Buffer.

 
  1. Analyze the expression of hematopoietic progenitor cell markers simultaneously by flow cytometry.
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Product Documents for Human Hematopoietic Progenitor Cell Multi-Color Flow Kit

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