EasySep® Human Whole Blood CD3 Positive Selection Kit

Recommended for:	The isolation of human CD3+ cells from whole blood by positive selection
Components:	• EasySep® Human Whole Blood CD3 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:	HIV, Immunology
Cell Isolation Product Type:	Reagents
Cell Source:	Buffy coat, Whole blood
Cell Type:	T cells
Popular Product Line:	EasySep, RoboSep
Selection Method:	Positive
Species:	Human

Procedures and instruction manuals:

•  PRODUCT INFORMATION SHEET - 18081, Lot# All:28948-PIS_1_3_0.pdf
•  PRODUCT INFORMATION SHEET - 18081RF, Lot# All:28948-PIS_1_3_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
•  VIDEO: HetaSep™: A Guide to Isolating Nucleated Cells from Whole Blood
•  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
•  VIDEO: The EasySep® MultiStand: Four Samples, Four Separations, One Pour-off
•  BROCHURE: Customized Cell Isolation: Flexible Cell Isolation Designed for you and your Research
•  SCIENTIFIC POSTER: Isolation of Highly Purified Subsets Directly from Whole Blood
•  SCIENTIFIC POSTER: A rapid automated method for the sequential isolation of CD19, CD3 and Myeloid cells from one tube of whole blood
•  VIDEO: RoboSep® Product Tour
•  VIDEO: Ficoll-Paque™ PLUS: A Guide to Isolating Mononuclear Cells from Whole Blood
•  VIDEO: RoboSep® Sneak Preview
•  BROCHURE: Fully Automated, High Purity Cell Isolation for Chimerism Labs
•  WALLCHART: Frequencies of Cell Types in Human Peripheral Blood Wallchart

MSDS:

•  18081C.1-MSDS.pdf
•  18150_18150H-MSDS.pdf

FAQS:

• 
  Q. HOW DOES THE SEPARATION WORK?
  
  A. Tiny magnetic particles are attached to cells. When placed in the EasySep® Magnet, the cells migrate to the wall of the tube.
• 
  Q. WHICH COLUMNS DO I USE?
  
  A. None, columns are not used in the EasySep^® procedure.
• 
  Q. CAN I ALTER THE SEPARATION TIME IN THE MAGNET?
  
  A. Yes, but this may affect separation performance.
• 
  Q. IN POSITIVE SELECTION CAN I DO MORE THAN 3 SEPARATIONS TO INCREASE PURITY?
  
  A. Yes, purity of targeted cells will increase with additional rounds of separations, however the recovery of targeted cells may decrease.
• 
  Q. ARE EASYSEP^® MAGNETIC NANOPARTICLES FACS-COMPATIBLE?
  
  A. Yes, the EasySep^® particles are flow cytometry-compatible, very uniform in size and ~5000X smaller than other commercially available magnetic beads used with column-free systems.
• 
  Q. CAN THE PARTICLE BE REMOVED AFTER ENRICHMENT?
  
  A. No, but they are so tiny, they do not interfere with downstream applications.
• 
  Q. CAN EASYSEP^® BE USED TO ISOLATE RARE CELLS?
  
  A. Yes, however we recommend a cell concentration of 2X10⁸ cells/ml and a minimum working volume of 100 µL. 2x10⁷ cells or fewer should be suspended in 100 µL of buffer.
• 
  Q. CAN EASYSEP^® BE USED FOR EITHER POSITIVE OR NEGATIVE SELECTION?
  
  A. Yes, we have products for both.
• 
  Q. HOW CAN I ANALYZE THE PURITY OF MY ENRICHED SAMPLE?
  
  A. Each Product Information Sheet has detailed staining information.
• 
  Q. HOW DOES THE BINDING OF THE NANOPARTICLE AFFECT THE FUNCTION OF CELLS? IS THE FUNCTION OF POSITIVELY SELECTED CELLS ALTERED BY THE BOUND PARTICLES?
  
  A. No, we have found that cells are not functionally altered.
• 
  Q. CAN EASYSEP^® SEPARATIONS BE AUTOMATED?
  
  A. Yes, click here for a link to RoboSep^®.

This product has been used in:

1. Endi Wang et al. Pseudo-Pelger-Hu??t anomaly induced by medications: a clinicopathologic study in comparison with myelodysplastic syndrome-related pseudo-Pelger-Hu??t anomaly.Am J Clin Pathol 135 (2) 291-303 (February 2011)

Product Name	Description	Catalog #
Ammonium Chloride Solution	Reagent for the Lysis of Red Blood Cells	07800
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
RoboSep® Red Blood Cell Lysis Buffer	Red Blood Cell Lysis Buffer	20110
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

FACS Histogram Results with EasySep® Human Whole Blood CD3 Positive Selection Kit

Starting with fresh peripheral blood, the CD3+ cell content of the selected fraction typically ranges from 98.2% - 99.8%. *Red blood cells were removed by lysis prior to flow cytometry.

Typical EasySep® Procedure for Human Cells