EasySep® Human Naïve CD8+ T Cell Enrichment Kit

Recommended for:	The isolation of human naive CD8+ T cells from fresh peripheral blood mononuclear cells by negative selection
Components:	• EasySep® Human Naïve CD8+ T Cell Enrichment Cocktial, 1.0 mL • EasySep® Human CD45RO Depletion Cocktial, 1.0 mL • EasySep® D2 Magnetic Particles, 2 x 1.0 mL (19158) • EasySep® D2 Magnetic Particles, 5 x 1.0 mL (19158RF)
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 • "Easy50" EasySep® Magnet (Catalog #18002), or • RoboSep® (Catalog #20000)
Product Type:	Cell isolation products
Area of Interest:	Immunology
Cell Isolation Product Type:	Reagents
Cell Source:	PBMC
Cell Type:	CD8+ T cells
Popular Product Line:	EasySep
Selection Method:	Negative
Species:	Human

Procedures and instruction manuals:

•  PRODUCT INFORMATION SHEET - 19158, Lot# All:28769-PIS_1_0_0.pdf
•  PRODUCT INFORMATION SHEET - 19158RF, Lot# All:28769-PIS_1_0_0.pdf

Educational resources:

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

MSDS:

•  19650-MSDS.pdf
•  19118C_19158C-MSDS.pdf
•  19126C_19146C-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.

Product Name	Description	Catalog #
CD8 Antibody, Clone SK1, FITC-Conjugated	Mouse Monoclonal Antibody to Human CD8 - FITC Conjugated	10405
Ficoll-Paque™ PLUS	Density Gradient Medium for the Isolation of Mononuclear Cells	07907
The Big Easy EasySep® Magnet	Immunomagnetic Column-Free Magnet	18001
RoboSep™	RoboSep™ - The Fully Automated Cell Separator	20000
CD8 Antibody, Clone SK1, PE-Conjugated	Mouse Monoclonal Antibody to Human CD8 - PE Conjugated	10505
EasySep® Magnet	Immunomagnetic Column-Free Magnet	18000

Start: 9.8% CD8+CD45RA+CCR7+CD45RO-CD56-CD57- of total peripheral blood mononuclear cells (PBMC)

Enriched: 91.4% CD8+CD45RA+CCR7+CD45RO-CD56-CD57- of enriched fraction

Starting with fresh mononuclear cells, the naïve CD8+ T cell content of the enriched fraction typically ranges from 85 - 92%.