EasySep® HLA CD3 Positive Selection Kit

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Immunomagnetic Positive Selection Kit

  • EasySep® box
  • Box top label for HLA CD3 Positive Selection Kit For labeling 10e9 total cells
  • Box top label for EasySep® HLA FCXM Blocking Solution Store at 15 - 25°C
EasySep® box
The EasySep® HLA CD3 Positive Selection Kit (Catalog #18051HLA) is designed to isolate CD3+ cells from fresh or previously frozen peripheral blood mononuclear cells by positive selection. Desired cells are targeted with Tetrameric Antibody Complexes recognizing CD3 and dextran-coated magnetic particles. This 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 CD3 antigen is expressed on all T cells and CD56+ NKT cells. Positively selected cells are immediately ready for flow cytometric crossmatch analysis (when used with EasySep® HLA FCXM Blocking Solution, included in kit), serology, or chimerism analysis.
Product Name Description Catalog # Size Price Quantity
EasySep® HLA CD3 Positive Selection Kit Column-free Immunomagnetic positive selection of human CD3+ cells (e.g. T cells) for HLA analysis 18051HLA For labeling up to 1 x 10e9 cells 545.00 USD      
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RoboSep® HLA CD3 Positive Selection Kit with Filter Tips EasySep® kit with RoboSep® buffer and RoboSep® filter tips 18051HLARF For labeling up to 1 x 10e9 cells 577.00 USD      
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Recommended for:
The isolation of human CD3+ cells from fresh or previously frozen peripheral blood mononuclear cells by positive selection
Components:
• EasySep® HLA CD3 Positive Selection Cocktail, 1.0 mL
• EasySep® Magnetic Particles, 1.0 mL
• EasySep® HLA FCXM Blocking Solution, 5 x 2.0 mL
Intended Use Statement: For Research Use Only. Not for Therapeutic or Diagnostic Use.
Equipment Required:
• EasySep® Magnet (Catalog #18000), or
• “The Big Easy” EasySep® Magnet (Catalog #18001), or
• RoboSep® (Catalog #20000)
Product Type: Cell isolation products
Area of Interest: Chimerism, HLA
Cell Isolation Product Type: Reagents
Cell Source: Other, PBMC
Cell Type: T 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 #

CD3 Antibody, Clone SK7, PE-Conjugated Mouse Monoclonal Antibody to Human CD3 - PE Conjugated 10502
CD3 Antibody, Clone SK7, FITC-Conjugated Mouse Monoclonal Antibody to CD3 - FITC Conjugated 10402
Ficoll-Paque™ PLUS Density Gradient Medium for the Isolation of Mononuclear Cells 07907
EasySep® Magnet Immunomagnetic Column-Free Magnet 18000
The Big Easy EasySep® Magnet Immunomagnetic Column-Free Magnet 18001

FACS Histogram Results with EasySep® HLA CD3+ Positive Selection Kit


FACS Histogram Results with EasySep® HLA CD3+ Positive Selection Kit
Starting with fresh peripheral blood mononuclear cells, the CD3+ cell content of the enriched fraction typically ranges from 94.6% - 99.6%.