EasySep® Mouse NK Cell Enrichment Kit

Recommended for:	The isolation of mouse NK cells from single cell suspensions of splenocytes by negative selection
Components:	• EasySep® Mouse NK Cell Enrichment Cocktail, 0.5 mL • EasySep® Biotin Selection Cocktail, 2 x 1.0 mL • EasySep® Magnetic Particles, 2 x 1.0 mL
Intended Use Statement:	For Research Use Only. Not for Therapeutic or Diagnostic Use.
Contains:
Equipment Required:	• EasySep® Magnet (Catalog #18000), or • “The Big Easy” EasySep® Magnet (Catalog #18001)
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:	Negative
Cell Isolation Product Type:	Reagents

Procedures and instruction manuals:

•  PRODUCT INFORMATION SHEET - 19755, Lot# All:28755-PIS_3_1_0.pdf

Educational resources:

•  VIDEO: The EasySep® Pour-off: A Simple and Effective Way to Isolate Cells
•  VIDEO: EasySep®: Powerful Immunomagnetic Isolation of Virtually Any Cell Type
•  BROCHURE: Cell Separation Equipment & Products
•  BROCHURE: Cell Separation for Autoimmunity Research
•  WALLCHART: Antigen Processing and Presentation Wallchart
•  BROCHURE: Ready Sep Go – Cell Separation In As Little As 25 Minutes
•  VIDEO: The EasySep® MultiStand: Four Samples, Four Separations, One Pour-off
•  BROCHURE: Customized Cell Isolation: Flexible Cell Isolation Designed for you and your Research
•  VIDEO: HetaSep™: A Guide to Isolating Nucleated Cells from Whole Blood
•  VIDEO: RoboSep® Product Tour
•  VIDEO: How to Isolate Mononuclear Cells from Whole Blood by Density Gradient Centrifugation
•  VIDEO: RoboSep® Sneak Preview
•  BROCHURE: Fast and Easy Isolation of Natural Killer Cells

MSDS:

•  19715C.2_19755C.2-MSDS.pdf
•  19250_19250H-MSDS.pdf

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. Manaf Bouchentouf et al. Induction of cardiac angiogenesis requires killer cell lectin-like receptor 1 and ??4??7 integrin expression by NK cells.J Immunol 185 (11) 7014-7025 (December 1, 2010)
2. Yuko Kawakami et al. Inhibition of NK cell activity by IL-17 allows vaccinia virus to induce severe skin lesions in a mouse model of eczema vaccinatumThe Journal of Experimental Medicine (May 25, 2009)

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 Profile Results with EasySep® Mouse NK Cell Enrichment Kit

Starting with C57BL6 mouse splenocytes, the CD49b+ CD3- cell content of the enriched fraction typically ranges from 75 - 87%.

Typical EasySep® Procedure for Mouse Cells