EasySep® Mouse panNK (CD49b) Positive Selection Kit

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Immunomagnetic Positive Selection Kit

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  • Box top label for Mouse panNK Positive Selection Kit For labeling 2 x 10e9 cells
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The EasySep® Mouse panNK (CD49b) Positive Selection Kit is designed to isolate CD49b+ cells from single cell suspensions of splenocytes or other tissues by positive selection. The cocktail contains a combination of PE-labeled antibodies that is directed against CD49b, which are then recognized by Tetrameric Antibody Complexes that are directed against PE and dextran. Labeled cells are bound to magnetic particles and separated using an EasySep® magnet without the use of columns. Cells of interest remain in the tube while unwanted cells are poured off.
Product Name Description Catalog # Size Price Quantity
EasySep® Mouse panNK (CD49b) Positive Selection Kit Immunomagnetic positive selection of CD49b+ cells (e.g. NK cells) from mouse splenocytes or other tissues 18755 For labeling up to 2 x 10e9 cells 587.00 USD      
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RoboSep® Mouse panNK (CD49b) Positive Selection Kit with Filter Tips EasySep® kit with RoboSep® buffer and RoboSep® filter tips 18755RF For labeling up to 2 x 10e9 cells 652.00 USD      
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Call us: +1.604.877.0713  
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Recommended for:
The isolation of mouse CD49b+ cells from single cell suspensions of splenocytes by positive selection
Components:
• EasySep® Mouse panNK (CD49b) PE Labeling Reagent, 1.0 mL
• EasySep® PE Selection Cocktail, 2 x 1.0 mL
• EasySep® Magnetic Particles, 1.0 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
Cell Type: NK cells
Popular Product Line: EasySep, RoboSep
Area of Interest: Immunology
Species: Mouse
Cell Source: Spleen, Other
Selection Method: Positive
Cell Isolation Product Type: Reagents

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.

This product has been used in:

  1. Alix E Seif et al. Long-term protection from syngeneic acute lymphoblastic leukemia by CpG ODN-mediated stimulation of innate and adaptive immune responses.Blood 114 (12) 2459-2466 (September 17, 2009)
  2. Elke Schneider et al. IL-33 activates unprimed murine basophils directly in vitro and induces their in vivo expansion indirectly by promoting hematopoietic growth factor production.J Immunol 183 (6) 3591-3597 (September 15, 2009)
  3. Mohammed S. Osman et al. Activating Ly-49 Receptors Regulate LFA-1-Mediated Adhesion by NK CellsJ Immunol 178 (3) 1261-1267 (February 1, 2007)
  4. Hortence Makui et al. Contribution of Hfe expression in macrophages to the regulation of hepatic hepcidin levels and iron loading.Blood 106 (6) 2189-2195 (September 15, 2005)
  5. Rima Koka et al. Cutting edge: murine dendritic cells require IL-15R alpha to prime NK cells.J Immunol 173 (6) 3594-3598 (September 15, 2004)

Product Name

Description

Catalog #
NK1.1 Antibody, Clone PK136, PE-Conjugated Mouse Monoclonal Antibody to Mouse NK1.1 - PE Conjugated 10827
NK1.1 Antibody, Clone PK136, FITC-Conjugated Mouse Monoclonal Antibody to Mouse NK1.1 - FITC Conjugated 10728
CD49b/panNK Antibody, Clone DX5, PE-Conjugated Rat Monoclonal Antibody to Mouse CD49b (panNK) - PE Conjugated 10826
CD49b/panNK Antibody, Clone DX5, FITC-Conjugated Rat Monoclonal Antibody to Mouse CD49b (panNK) - FITC Conjugated 10726
EasySep® Magnet Immunomagnetic Column-Free Magnet 18000
The Big Easy EasySep® Magnet Immunomagnetic Column-Free Magnet 18001
RoboSep™ RoboSep™ - The Fully Automated Cell Separator 20000

FACS Histogram Results with EasySep® Mouse panNK (CD49b) Selection Kit


FACS Histogram Results with EasySep® Mouse panNK (CD49b) Selection Kit
Starting with mouse splenocytes, the CD49b+ (DX5) cell content of the selected cells typically ranges from 88 - 97%.


Typical EasySep® Procedure for Human Cells


Typical EasySep® Procedure for Human Cells



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