EasySep™ Human Neutrophil Enrichment Kit

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Immunomagnetic Negative Selection Kit

  • EasySep™ Box
  • Box top label for Human Neutrophil Enrichment Kit For labeling 10e9 total cells
EasySep™ Box
The EasySep™ Human Neutrophil Enrichment Kit is designed to isolate neutrophils from a polymorphonuclear cell-rich fraction of peripheral blood by negative selection. Unwanted cells are targeted for removal with Tetrameric Antibody Complexes recognizing CD2, CD3, CD9, CD19, CD36, CD56, glycophorin A and dextran-coated magnetic particles. Labeled cells are separated using an EasySep™ magnet without the use of columns. Desired cells are poured off into a new tube. The enrichment kit is compatible with cells prepared using HetaSep™ (Catalog #07906) sedimentation or red blood cell lysis.
Product Name Description Catalog # Size Price Quantity
EasySep™ Human Neutrophil Enrichment Kit Immunomagnetic isolation of untouched human neutrophils 19257 For labeling up to 1 x 10e9 cells 881.00 USD      
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RoboSep™ Human Neutrophil Enrichment Kit with Filter Tips EasySep™ kit with RoboSep™ buffer and RoboSep™ filter tips 19257RF For labeling up to 1 x 10e9 cells 916.00 USD      
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Call Us: +1.604.877.0713  
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Product Type: Cell isolation products
Recommended For:
The isolation of neutrophils from a polymorphonuclear cell-rich fraction by negative selection
Components:
• EasySep™ Human Neutrophil Enrichment Cocktail, 1.0 mL
• EasySep™ Magnetic Particles, 3 x 1.0 mL
Equipment Required:
Area of Interest: Immunology
Cell Isolation Product Type: Reagents
Cell Source: PMNC
Cell Type: Granulocytes & subsets
Popular Product Line: EasySep™, RoboSep™
Selection Method: Negative
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™ Human Neutrophil Enrichment Kit All Product Information Sheet
RoboSep™ Human Neutrophil Enrichment Kit with Filter Tips All Product Information Sheet
 

Educational resources:

MSDS:

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.
 

This product has been used in:

  1. Nicola Tamassia et al. Cutting edge: An inactive chromatin configuration at the IL-10 locus in human neutrophils.J Immunol 190 (5) 1921-5 (March 1, 2013)
  2. Marie-Ming Aynaud et al. Human Tribbles 3 protects nuclear DNA from cytidine deamination by APOBEC3A.J Biol Chem 287 (46) 39182-92 (November 9, 2012)
  3. Claudio Costantini et al. Neutrophil activation and survival are modulated by interaction with NK cells.Int Immunol 22 (10) 827-838 (October 2010)
  4. Hyun Sik Jun et al. Lack of glucose recycling between endoplasmic reticulum and cytoplasm underlies cellular dysfunction in glucose-6-phosphatase-beta-deficient neutrophils in a congenital neutropenia syndrome.Blood 116 (15) 2783-2792 (October 14, 2010)
  5. Sven Brode et al. Interleukin-5 inhibits glucocorticoid-mediated apoptosis in human eosinophils.Thorax (August 30, 2010)
  6. Andrew S Cowburn et al. GM-CSF causes a Paradoxical Increase in the BH3-only Pro-Apoptotic Protien BIM in Human Neutrophils.Am J Respir Cell Mol Biol (August 12, 2010)
  7. Rapha??lle Romieu-Mourez et al. Cytokine modulation of TLR expression and activation in mesenchymal stromal cells leads to a proinflammatory phenotype.J Immunol 182 (12) 7963-7973 (June 15, 2009)

Product Name

Description

Catalog #

HetaSep™ For Isolating Human Nucleated Cells from Peripheral Blood 07806
EasySep™ Magnet Immunomagnetic Column-Free Magnet 18000
"The Big Easy" EasySep™ Magnet Immunomagnetic Column-Free Magnet 18001
RoboSep™-S RoboSep™-S - The Fully Automated Cell Separator 21000
Anti-Human CD32 Antibody, Clone IV.3 Mouse Monoclonal Antibody Against Human CD32 60012
Anti-Human CD45 Antibody, Clone HI30 Mouse Monoclonal Antibody Against Human, Chimpanzee CD45 60018
Anti-Dextran Antibody, Clone DX1 Mouse Monoclonal Antibody Against Dextran 60026

FACS Profile Results Using EasySep™ Human Neutrophil Enrichment Kit


FACS Profile Results Using EasySep™ Human Neutrophil Enrichment Kit
Starting with fresh HetaSep™-treated blood, the neutrophil content of the enriched fraction typically ranges from 98% - 99%.


Typical EasySep™ Procedure for Human Cells


Typical EasySep™ Procedure for Human Cells



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