EasySep® Human Whole Blood CD34 Positive Selection Kit

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Immunomagnetic Positive Selection Kit

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  • Box top label for Human Whole Blood/Buffy Coat CD34 Positive Selection Kit For use with Whole Blood or Buffy Coat
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The EasySep® Human Whole Blood CD34 Positive Selection Kit is designed to isolate CD34+ cells from fresh whole blood or freshly prepared buffy coat by positive selection. Desired cells are targeted with Tetrameric Antibody Complexes recognizing CD34 and dextran-coated magnetic particles. The cocktail also contains an antibody to human Fc receptor to minimize nonspecific binding. Labeled cells are separated using an EasySep® magnet without the use of columns. Cells of interest remain in the tube while unwanted cells are poured off. The CD34 antigen is expressed on hematopoietic stem and progenitor cells. If isolating CD34+ cells from fresh cord blod, we recommend using the EasySep® Human Cord Blood CD34 Positive Selection Kit (Catalog #18096). If isolating CD34+ cells from any other sample tyep, we recommend using the EasySep® Human CD34 Positive Selection Kit (Catalog #18056).
Product Name Description Catalog # Size Price Quantity
EasySep® Human Whole Blood CD34 Positive Selection Kit Immunomagnetic positive selection of human CD34+ cells (e.g. hematopoietic stem/progenitor cells) from whole blood or buffy coat 18086 For labeling 75 mL of whole blood (37 mL buffy coat) 507.00 USD      
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RoboSep® Human Whole Blood CD34 Positive Selection Kit with Filter Tips EasySep® kit with RoboSep® buffer and RoboSep® filter tips 18086RF For labeling 75 mL of whole blood (37 mL buffy coat) 568.00 USD      
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Recommended for:
The isolation of human CD34+ cells from fresh whole blood or freshly prepared buffy coat by positive selection
Components:
• EasySep® Human Whole Blood CD34 Positive Selection Cocktail, 3 x 1.0 mL
• EasySep® Magnetic Particles, 3 x 1.0 mL
• EasySep® RBC Lysis Buffer 10X Concentrate, 10 mL
Intended Use Statement: For Research Use Only. Not for Therapeutic or Diagnostic Use.
Equipment Required:
• “The Big Easy” EasySep® Magnet (Catalog #18001), or
• RoboSep® (Catalog #20000)
Product Type: Cell isolation products
Area of Interest: Chimerism, Hematopoietic stem cell research, HLA, Stem cell biology
Cell Isolation Product Type: Reagents
Cell Source: Buffy coat, Whole blood
Cell Type: Hematopoietic cells, Hematopoietic stem & progenitor cells
Popular Product Line: EasySep, RoboSep
Selection Method: Positive
Species: Human

Procedures and instruction manuals:

Educational resources:

MSDS:

FAQS:


  • HOW EASYSEP™ WORKS

    • Q. CAN EASYSEP™ BE USED FOR EITHER POSITIVE OR NEGATIVE SELECTION?
      A. Yes. The EasySep™ kits use either a negative selection approach by targeting and removing unwanted cells or a positive selection approach targeting desired cells. Depletion kits are also available for the removal of cells with a specific undesired marker (e.g. GlyA).
    • Q. HOW DOES THE SEPARATION WORK?
      A. Magnetic particles are crosslinked to cells using Tetrameric Antibody Complexes (TAC). When placed in the EasySep™ Magnet, labeled cells migrate to the wall of the tube. The unlabeled cells are then poured off into a separate fraction.
    • Q. WHICH COLUMNS DO I USE?
      A. The EasySep™ procedure is column-free. That's right - no columns!
    • Q. HOW CAN I ANALYZE THE PURITY OF MY ENRICHED SAMPLE?
      A. The Product Information Sheet provided with each EasySep™ kit contains detailed staining information.
    • Q. CAN EASYSEP™ SEPARATIONS BE AUTOMATED?
      A. Yes. RoboSep™, the fully automated cell separator, automates all EasySep™ labeling and cell separation steps.
    • Q. CAN EASYSEP™ BE USED TO ISOLATE RARE CELLS?
      A. Yes. We recommend a cell concentration of 2x108 cells/mL and a minimum working volume of 100 µL. Samples containing 2x107 cells or fewer should be suspended in 100 µL of buffer.

    EASYSEP™ MAGNETIC PARTICLES

    • Q. ARE THE EASYSEP™ MAGNETIC PARTICLES FACS-COMPATIBLE?
      A. Yes, the EasySep™ particles are flow cytometry-compatible, as they are very uniform in size and about 5000X smaller than other commercially available magnetic beads used with column-free systems.
    • Q. CAN THE EASYSEP™ MAGNETIC PARTICLES BE REMOVED AFTER ENRICHMENT?
      A. No, but due to the small size of these particles, they will not interfere with downstream applications.
    • Q. HOW DOES THE BINDING OF THE EASYSEP™ MAGNETIC PARTICLE AFFECT THE CELLS? IS THE FUNCTION OF POSITIVELY SELECTED CELLS ALTERED BY THE BOUND PARTICLES?
      A. Hundreds of publications have used cells selected with EasySep™ positive selection kits for functional studies. Our in-house experiments also confirm that selected cells are not functionally altered by the EasySep™ magnetic particles.

      If particle binding is a key concern, we offer two options for negative selection. The EasySep™ negative selection kits can isolate untouched cells with comparable purities, while RosetteSep™ can isolate untouched cells directly from whole blood without using particles or magnets.

    MODIFYING THE EASYSEP™ PROTOCOL

    • Q. CAN I ALTER THE SEPARATION TIME IN THE MAGNET?
      A. Yes; however, this may impact the kit's performance. The provided EasySep™ protocols have already been optimized to balance purity, recovery and time spent on the isolation.
    • Q. FOR POSITIVE SELECTION, CAN I PERFORM MORE THAN 3 SEPARATIONS TO INCREASE PURITY?
      A. Yes, the purity of targeted cells will increase with additional rounds of separations; however, cell recovery will decrease.

Product Name

Description

Catalog #
CD34 Antibody, Clone 8G12, PE-Conjugated Mouse Monoclonal Antibody to Human CD34 - PE Conjugated 10513
CD34 Antibody, Clone 8G12, FITC-Conjugated Mouse Monoclonal Antibody to Human CD34 - FITC Conjugated 10413
CD34 Antibody, Clone 8G12, APC-Conjugated Mouse Monoclonal Antibody to Human CD34 - PE Conjugated 10613
Ficoll-Paque™ PLUS Density Gradient Medium for the Isolation of Mononuclear Cells 07907
RoboSep® Red Blood Cell Lysis Buffer Red Blood Cell Lysis Buffer 20110
Ammonium Chloride Solution Reagent for the Lysis of Red Blood Cells 07800
The Big Easy EasySep® Magnet Immunomagnetic Column-Free Magnet 18001
RoboSep™ RoboSep™ - The Fully Automated Cell Separator 20000
EasySep® Magnet Immunomagnetic Column-Free Magnet 18000
MethoCult® H4034 Optimum Methylcellulose-Based Medium with Recombinant Cytokines and EPO for Human Cells 04034
MethoCult® H4435 Enriched Methylcellulose Medium with Recombinant Cytokines and EPO for Human Cells 04435
MethoCult® H4434 Classic Methylcellulose-Based Medium with Recombinant Cytokines for Human Cells 04434

FACS Profile Results with EasySep® Human Buffy Coat/Whole Blood CD34 Positive Selection Kit


FACS Profile Results with EasySep® Human Buffy Coat/Whole Blood CD34 Positive Selection Kit
Starting with buffy coat, the CD34+ cell content of the enriched fraction typically ranges from 51.7% - 90.3% (reported as a percentage of viable CD45+ cells).


Typical EasySep® Procedure for Human Cells


Typical EasySep® Procedure for Human Cells



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