EasySep® Human Plasmacytoid DC Enrichment Kit

Recommended for:	The isolation of human plasmacytoid dendritic cells (pDC) from fresh peripheral blood mononuclear cells by negative selection
Components:	• EasySep® Human Plasmacytoid DC Enrichment Cocktail, 2.0 mL • EasySep® Magnetic Particles, 8 x 1.0 mL • Anti-Human CD32 (FcγRII) Blocker, 1.6 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:	HIV, Immunology
Cell Isolation Product Type:	Reagents
Cell Source:	PBMC
Cell Type:	Dendritic cells
Popular Product Line:	EasySep, RoboSep
Selection Method:	Negative
Species:	Human

Procedures and instruction manuals:

•  PRODUCT INFORMATION SHEET - 19062, Lot# 10D3434 or greater:29149-PIS_2_1_0.pdf
•  PRODUCT INFORMATION SHEET - 19062, Lot# 10D3434 or greater:29149-Easy50-PIS_1_0_0.pdf
•  PRODUCT INFORMATION SHEET - 19062RF, Lot# All:29149-PIS_2_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
•  VIDEO: HetaSep™: A Guide to Isolating Nucleated Cells from Whole Blood
•  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
•  SCIENTIFIC POSTER: A Simple One-Step Method for isolating Highly Purified Plasmacytoid Dendritic Cells from Human Peripheral Blood
•  VIDEO: RoboSep® Product Tour
•  VIDEO: Ficoll-Paque™ PLUS: A Guide to Isolating Mononuclear Cells from Whole Blood
•  VIDEO: RoboSep® Sneak Preview
•  WALLCHART: Frequencies of Cell Types in Human Peripheral Blood Wallchart

MSDS:

•  19022C_19062C-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. 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. CAN EASYSEP^® SEPARATIONS BE AUTOMATED?
  
  A. Yes, click here for a link to RoboSep^®.

This product has been used in:

1. Nicholas J Megjugorac et al. IL-4 enhances IFN-lambda1 (IL-29) production by plasmacytoid DCs via monocyte secretion of IL-1Ra.Blood 115 (21) 4185-4190 (May 27, 2010)
2. Nicholas J Megjugorac et al. Modulation of human plasmacytoid DC function by IFN-lambda1 (IL-29).J Leukoc Biol 86 (6) 1359-1363 (December 2009)

Product Name	Description	Catalog #
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
RoboSep®	RoboSep® - The Fully Automated Cell Separator	20000

Typical EasySep® Human pDC Enrichment Profile

Starting with 0.2 - 0.9% pDC in PBMC, the pDC content of the enriched fraction typically ranges from 87 - 97% purity based on the pDC phenotype of Lineage (CD3, CD14, CD16, CD19, CD20, CD34, CD56) negative, HLA-DR positive, and CD304 (BDCA-4) positive.

Typical EasySep® Procedure for Human Cells

FACS Purity Data from pDC Kit User

FACS enrichment plots from Dr. Stuart R. McGregor Dallas of Princeton University. Prior to enrichment, the percentage of pDCs in total PBMC is approximately 0.1% (upper row), however following enrichment, a population of pDCs in excess of 97% pure can be obtained (lower row). pDCs identified using surface marker staining for CD303 and HLA-DR. Data originally posted as part of a product review in Biocompare.