Human Hematopoietic Progenitor Cell Multi-Color Flow Kit

Name: Human Hematopoietic Progenitor Cell Multi-Color Flow Kit
Brand: R&D Systems
SKU: FMC019

Catalog # FMC019 | R&D Systems, Inc. a Bio-Techne Brand

Contains conjugated antibodies to CD34-APC (Clone QBEnd10), CD38-PerCP (Clone 240742), CD11b-Fluorescein (Clone 238446), SCF R-PE (Clone 74233)

Key Product Details

Setting the standard in quality research reagents

Summary for Human Hematopoietic Progenitor Cell Multi-Color Flow Kit

Kit Summary

To verify hematopoietic progenitor cell multipotency using 4-color flow cytometry.

Why is it Important to Verify Hematopoietic Progenitor Cell Identity using Multiple Established Markers?

Hematopoietic stem cells are multipotent, self-renewing progenitor cells from which all differentiated blood cell types arise during the process of hematopoiesis. Using multiple markers to characterize hematopoietic progenitors increases confidence in cell identity by clearly and consistently defining your target cell population. See Details

In addition, the unique combination of negative and positive markers included in this kit decreases experimental variation in hematopoietic progenitor cell identification, resulting in an optimized starting cell population for your research.

Uses a combination of positive and negative cell-surface markers to verify progenitor cell identity
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

This flow cytometry kit contains four fluorochrome-conjugated primary antibodies and corresponding isotype controls that can be used for single-step staining of human hematopoietic progenitor cells. This kit includes reagents sufficient for 25 tests. See Details

Staining Buffer (100 mL)

Positive Markers:

APC-conjugated Mouse Anti-Human CD34 (Clone QBEnd10; Mouse IgG₁)
PE-conjugated Mouse Anti-Human SCF R/CD117 (Clone 47233; Mouse IgG₁)

Negative Markers:

PerCP-conjugated Mouse Anti-Human CD38 (Clone 240742; Mouse IgG_2A)
CFS-conjugated Mouse Anti-Human CD11b (Clone 238446,IgG_2B)

Positive Isotype Controls

APC-conjugated Mouse IgG₁ Isotype Control (Clone 11711)
PE-conjugated Mouse IgG₁ Isotype Control (Clone 11711)

Negative Isotype Controls

PerCP-conjugated Mouse IgG_2A Isotype Control (Clone 20102)
CFS-conjugated Mouse IgG_2B Isotype Control (Clone 133303)

HSC Differentiation &Lineage Marker Schematic View Interactive Pathway

The definitive hematopoietic system is made up of all adult blood cell types including megakaryocytes, erythrocytes, and cells of the myeloid and lymphoid lineages. All of these cells are derived from multipotent hematopoietic stem cells (HSCs) through a succession of precursors with progressively limited potential. Hematopoietic stem cells are tissue-specific stem cells that exhibit remarkable self-renewal capacity and are responsible for the life-long maintenance of the hematopoietic system. HSCs are rare cells that reside in adult bone marrow where hematopoiesis is continuously taking place. They can also be found in cord blood, fetal liver, adult spleen, and peripheral blood. R&D Systems offers several products for studying hematopoietic lineage cells including serum-free media, lineage depletion antibodies and kits, and reagents for performing colony forming cell (CFC) assays.

Product Specifications for Human Hematopoietic Progenitor Cell Multi-Color Flow Kit

Species

Human

Scientific Data Examples for Human Hematopoietic Progenitor Cell Multi-Color Flow Kit

	Identification of Human Hematopoietic Progenitor Cells using Multi-Color Flow Cytometry. Human umbilical cord blood cells were stained using reagents supplied in this kit. Cells were simultaneously analyzed for the expression of multipotency markers. Cells negative for CD11b and negative/low for CD38 (boxed area in A) were gated and assessed for positive expression of CD34 and SCF R/CD117 (upper right quadrant in B). Quadrants were set based on isotype controls.
	Identification of Human Hematopoietic Progenitor Cells using Multi-Color Flow Cytometry. Human umbilical cord blood cells were stained with the indicated antibodies included in this kit. Representative expression of each analyte in the total umbilical cord blood cell population (filled histogram) over the isotype control (open histogram).

Preparation & Storage

Shipping Conditions	The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Storage	Store the unopened product at 2 - 8 °C. Do not use past expiration date.

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

View Graphic Procedure

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 IgG_2A)
CFS-conjugated Mouse Anti-Human CD11b (Clone 238446,IgG_2B)
APC-conjugated Mouse IgG₁ Isotype Control (Clone 11711)
PE-conjugated Mouse IgG₁ Isotype Control (Clone 11711)
PerCP-conjugated Mouse IgG_2A Isotype Control (Clone 20102)
CFS-conjugated Mouse IgG_2B 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

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

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 10⁵ 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 hematopoietic progenitor cell markers simultaneously by flow cytometry.

Citations for Human Hematopoietic Progenitor Cell Multi-Color Flow Kit

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