EasySep® hESC/hiPSC SSEA-4 Positive Selection Kit

Procedures and instruction manuals:

•  PRODUCT INFORMATION SHEET - 18165, Lot# All:29974-PIS_2_0_0.pdf

Educational resources:

•  WALLCHART: Pluripotent Stem Cell Biology Wallchart
•  BROCHURE: Standardized Solutions for hESC & hiPSC Research
•  VIDEO: The EasySep® MultiStand: Four Samples, Four Separations, One Pour-off
•  VIDEO: EasySep®: Powerful Immunomagnetic Isolation of Virtually Any Cell Type
•  VIDEO: The EasySep® Pour-off: A Simple and Effective Way to Isolate Cells

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.

Testimonials:

• I have been using this system for the past year on a monthly basis and have found this technique to be a reproducible and reliable way of isolating populations of pluripotent stem cells. The main advantages of this system are the efficiency of the isolation procedure with a high purity (98-99%) achieved post-isolation, as identified through FACS analysis. Also, the passive nature of this procedure allows isolated cells to be cultured without detrimentally affecting cell viability. Since the antibody-magnetic bead complex remains attached to the cells post-isolation, downstream applications could be affected; however, it did not appear to affect cell viability of hESCs post-isolation.
  
  Overall, this system is an excellent way of isolating pluripotent stem cells based on the expression of the SSEA4 antigen. Read More

  - Fiona Lewis

FACS Histogram Results Using EasySep® hESC/hiPSC SSEA-4 Positive Selection Kit

Typical EasySep® Procedure for Human Cells