EasySep™ hESC/hiPSC SSEA-4 Positive Selection Kit

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Immunomagnetic Positive Selection Kit.

  • EasySep™ Box
  • hESC/hiPSC SSEA-4 Positive Selection Kit Label
EasySep™ Box
The EasySep™ hESC/hiPSC SSEA-4 Positive Selection Kit is designed for the positive selection of undifferentiated human embryonic stem and induced pluripotent stem cells. The cocktail contains antibodies directed against SSEA-4, which are then linked to anti-dextran antibodies through a unique system. Labeled cells are bound to magnetic particles and separated using an EasySep™ magnet without the use of columns. Cells of interest remain in the tube while unwanted cells are poured off.
Product Name Description Catalog # Size Price Quantity
EasySep™ hESC/hiPSC SSEA-4 Positive Selection Kit Immunomagnetic positive selection of hESC/hiPSC SSEA-4+ cells 18165 For labeling up to 1 x 10e9 total cells 666.00 USD      
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Clone Name: MC-813-70
Immunogen: Human embryonal carcinoma cell line 2102 Ep
Isotype: IgG3, kappa
Product Type: Cell isolation products
Recommended For:
The isolation of undifferentiated human embryonic stem and induced pluripotent stem cells (SSEA-4+ cells) by positive selection.
Components:
• EasySep™ hESC/hiPSC SSEA-4 Selection Cocktail, 1.0 mL
• EasySep™ Magnetic Particles,  1.0 mL
Equipment Required:
Area of Interest: Embryonic stem cell & iPS cell research
Cell Isolation Product Type: Reagents
Cell Source: Other
Cell Type: Embryonic stem cells & iPS cells
Popular Product Line: EasySep™
Selection Method: Positive
Species: Human
Intended Use Statement: FOR RESEARCH USE ONLY. NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES.
ISO Statement: STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485 MEDICAL DEVICE STANDARDS

Protocols and Manuals

Current documents
Product NameLot# ?Document TypeDownload
EasySep™ hESC/hiPSC SSEA-4 Positive Selection Kit All Product Information Sheet
 

Educational resources:

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.
    - Fiona Lewis

Product Name

Description

Catalog #

EasySep™ hESC-Derived CD34 Positive Selection Kit Immunomagnetic Positive Selection Kit 18167
TeSR™2 Animal protein-free, defined, feeder-independent medium for maintenance of undifferentiated human ES and iPS cells 05860
mTeSR™1 Defined, Feeder-Free Maintenance Medium for Human ES and iPS Cells 05850
Anti-Human SSEA-4 Antibody, Clone MC-813-70 Mouse Monoclonal Antibody Against Human, Mouse, Rat SSEA-4 60062
Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R Mouse Monoclonal Antibody Against Human, Rhesus, Rabbit TRA-1-60 (Podocalyxin) 60064
EasySep™ Magnet Immunomagnetic Column-Free Magnet 18000

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


FACS Histogram Results Using EasySep™ hESC/hiPSC SSEA-4 Positive Selection Kit
Starting with a single cell suspension containing approximately 10% hESCs or hiPSCs, the SSEA-4+ cell content of the enriched fraction typically ranges from 85% to 95%.


Typical EasySep™ Procedure for Human Cells


Typical EasySep™ Procedure for Human Cells



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