Complete Kit for Human CD4+CD25+ T Cells

Procedures and instruction manuals:

•  PRODUCT INFORMATION SHEET - 15862, Lot# All:28571-PIS_1_2_0.pdf
•  PRODUCT INFORMATION SHEET - 15862, Lot# All:28985-PIS_2_3_1.pdf
•  PRODUCT INFORMATION SHEET - 15862RF, Lot# All:28571-PIS_1_2_0.pdf
•  PRODUCT INFORMATION SHEET - 15862RF, Lot# All:28985-PIS_2_3_1.pdf

Educational resources:

•  VIDEO: EasySep®: Powerful Immunomagnetic Isolation of Virtually Any Cell Type
•  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
•  VIDEO: RosetteSep® Introduction
•  VIDEO: RosetteSep®: A Guide to Isolating NK Cells Directly from Whole Blood
•  BROCHURE: One-Step Cell Enrichment Directly From Whole Blood
•  BROCHURE: RoboSep®: Solutions for HIV Research
•  BROCHURE: Simple and Effective Isolation of Human Regulatory T Cells
•  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
•  SCIENTIFIC POSTER: A simple and versatile method for the isolation of human regulatory T cells from peripheral blood samples ranging from 5 to 500 mL
•  SCIENTIFIC POSTER: A Simple Two-Step Method for the Isolation of Human CD4+CD25+bright/FOXP3+ Regulatory T Cells Directly from Whole Blood
•  SCIENTIFIC POSTER: Rapid isolation of CD4+CD25+ from whole blood
•  VIDEO: RosetteSep® for HLA Analysis
•  BROCHURE: Cell isolation products for regulatory T cells
•  TECH BULLETIN: Techinical Note: Human Regulatory T Cells (Catalog #29147)
•  WALLCHART: Frequencies of Cell Types in Human Peripheral Blood Wallchart

MSDS:

•  15012C.1_15022C.1-MSDS.pdf
•  18150_18150H-MSDS.pdf
•  18231C.2-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 TO ISOLATE CELLS FROM WHOLE BLOOD?
  
  A. Yes.
• 
  Q. CAN EASYSEP^® BE USED TO DEPLETE UNWANTED CELLS?
  
  A. Yes, we have depletion products available.
• 
  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^® BE USED TO ISOLATE CELLS FROM RAT, NON-HUMAN PRIMATE & OTHER SPECIES?
  
  A. Yes, provided an applicable antibody is available.
• 
  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 BSA BE USED IN PLACE OF FBS?
  
  A. Yes, 0.5% BSA can be substituted.
• 
  Q. CAN EASYSEP^® SEPARATIONS BE AUTOMATED?
  
  A. Yes, click here for a link to RoboSep^®.
• 
  Q. WHAT IS ROSETTESEP^®?
  
  A. RosetteSep^® is a rapid cell separation procedure for the isolation of purified cells directly from whole blood without columns or magnets.
• 
  Q. HOW DOES ROSETTESEP^® WORK?
  
  A. The antibody cocktail crosslinks unwanted cells to red blood cells (RBCs), forming rosettes. The unwanted cells then pellet with the free RBCs when centrifuged over a buoyant density medium.
• 
  Q. WHICH CELL SAMPLES CAN ROSETTESEP^® BE USED WITH?
  
  A. RosetteSep^® can be used with leukapheres, bone marrow samples or buffy coat, as long as: the concentration of cells does not exceed 5 x 10⁷ per mL (can dilute if necessary); and there are at least 20 RBCs for every nucleated cell (RBCs can be added if necessary).  For a full procedure click here.
• 
  Q. CAN ROSETTESEP^® BE USED WITH PREVIOUSLY FROZEN OR CULTURED CELLS?
  
  A. Yes. Cells should be re-suspended at 2 – 5 x 10⁷ cells / mL in PBS + 2% FBS. Fresh whole blood should be added at 250 µL per mL of sample, as a source of red cells.
• 
  Q. WHICH ANTICOAGULANT SHOULD BE USED WITH ROSETTESEP^®?
  
  A. Peripheral blood should be collected in heparinized Vaccutainers. Cord blood should be collected in ACD.
• 
  Q. SHOULD THE ANTICOAGULANT BE WASHED OFF BEFORE USING ROSETTESEP^®?
  
  A. No, the antibody cocktail can be added directly to the sample.
• 
  Q. CAN ROSETTESEP^® BE USED TO ENRICH PROGENITORS FROM CORD BLOOD?
  
  A. Yes. Sometimes cord blood contains immature nucleated red cells that have a lower density than mature RBCs. These immature red cells do not pellet over Ficoll^TM, which can lead to a higher RBC contamination than peripheral blood separations.
• 
  Q. DOES ROSETTESEP^® WORK WITH MOUSE CELLS?
  
  A. No, but we have developed EasySep^®, a magnetic-based separation for the isolation of mouse cells.

This product has been used in:

1. Rajendra Pahwa et al. Isolation and expansion of human natural T regulatory cells for cellular therapy.J Immunol Methods (October 23, 2010)
2. Xiao-Nong Wang et al. Regulatory T-cell suppression of CD8+ T-cell-mediated graft-versus-host reaction requires their presence during priming.Transplantation 88 (2) 188-197 (July 27, 2009)

FACS Profile Results With EasySep® Human CD25bright Cell Positive Selection Kit (following RosetteSep® Human CD4+ T Cell Enrichment)

RosetteSep® Procedure

Typical EasySep® Procedure for Human Cells