EasySep™ Human Monocyte Enrichment Kit without CD16 Depletion

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Immunomagnetic Negative Selection Kit

  • EasySep™ Box
  • Box top label for Human Monocyte Enrichment Kit without CD16 Depletion For labeling 10e9 cells
EasySep™ Box
The EasySep™ Human Monocyte Enrichment Kit without CD16 Depletion is designed to isolate monocytes and CD14+CD16+ monocytes from fresh or previously frozen peripheral blood mononuclear cells by negative selection. The CD14+CD16+ subset of monocytes (~10% in blood of healthy individuals) has characteristics of tissue macrophages and expands greatly in acute and chronic inflammatory disease. Unwanted cells are targeted for removal with Tetrameric Antibody Complexes recognizing CD2, CD3, CD19, CD20, CD56, CD66b, CD123, glycophorin A and dextran-coated magnetic particles. The cocktail also contains and antibody to human Fc receptor to minimize nonspecific binding. The labeled cells are separated using an EasySep™ magnet without the use of columns. Desired cells are poured off into a new tube. For applications in which removal of all CD16+ cells is desired, we recommend the EasySep™ Human Monocyte Enrichment Kit (Catalog #19059), which contains anti-CD16.
 
See below, "Equipment Required", for compatible magnets platforms.
Product Name Description Catalog # Size Price Quantity
EasySep™ Human Monocyte Enrichment Kit without CD16 Depletion Immunomagnetic isolation of untouched human monocytes 19058 For labeling up to 1 x 10e9 cells 685.00 USD      
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RoboSep™ Human Monocyte Enrichment Kit without CD16 Depletion with Filter Tips EasySep™ kit with RoboSep™ buffer and RoboSep™ filter tips 19058RF For labeling up to 1 x 10e9 cells 721.00 USD      
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Product Type: Cell isolation products
Recommended For:
The isolation of human monocytes and CD14+CD16+ monocytes from fresh or previously frozen peripheral blood mononuclear cells by negative selection
Components:
• EasySep™ Human Monocyte Enrichment Cocktail without CD16 Depletion, 1.0 mL
• EasySep™ Magnetic Particles, 1.0 mL
Equipment Required:
Area of Interest: HIV, Immunology
Cell Isolation Product Type: Reagents
Cell Source: Leukapheresis, PBMC
Cell Type: Monocytes
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 Monocyte Enrichment Kit without CD16 Depletion All Product Information Sheet
EasySep™ Human Monocyte Enrichment Kit without CD16 Depletion All Product Information Sheet - Easy50
EasySep™ Human Monocyte Enrichment Kit without CD16 Depletion All Product Information Sheet - EasyPlate
RoboSep™ Human Monocyte Enrichment Kit without CD16 Depletion 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. Marc Daigneault et al. Monocytes regulate the mechanism of T-cell death by inducing Fas-mediated apoptosis during bacterial infection.PLoS Pathog 8 (7) e1002814 (July 19, 2012)
  2. Carolina Garc??a-de-Alba et al. Expression of Matrix Metalloproteases by Fibrocytes. Possible Role in Migration and Homing.Am J Respir Crit Care Med (July 9, 2010)
  3. Shanshan Lin et al. HIV infection upregulates Caveolin 1 (Cav-1) expression to restrict virus production.J Virol (July 7, 2010)
  4. 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)
  5. Martin Pelletier et al. Evidence for a cross-talk between human neutrophils and Th17 cells.Blood 115 (2) 335-343 (January 14, 2010)
  6. Ryan A Wilcox et al. Monocytes promote tumor cell survival in T-cell lymphoproliferative disorders and are impaired in their ability to differentiate into mature dendritic cells.Blood 114 (14) 2936-2944 (October 1, 2009)

Product Name

Description

Catalog #

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EasySep™ Magnet Immunomagnetic Column-Free Magnet 18000
"The Big Easy" EasySep™ Magnet Immunomagnetic Column-Free Magnet 18001
Easy 50 EasySep™ Magnet Immunomagnetic Column-Free Magnet 18002
EasySep™ Human Monocyte Enrichment Kit Immunomagnetic Negative Selection Kit 19059
RoboSep™-S RoboSep™-S - The Fully Automated Cell Separator 21000
Anti-Human CD14 Antibody, Clone M5E2 Mouse Monoclonal Antibody Against Human, Rhesus, Cynomolgus CD14 60004
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 Histogram Results Using EasySep™ Human Monocyte Enrichment KitWithout CD16 Depletion


FACS Histogram Results Using EasySep™ Human Monocyte Enrichment KitWithout CD16 Depletion
Starting with freshly prepared peripheral blood mononuclear cells, the CD14+ cell content of the enriched fraction typically ranges from 73% - 81%. Slightly lower CD14+ cell purities may be obtained from samples that contain a large number of CD16+ cells.


Typical EasySep™ Procedure for Human Cells


Typical EasySep™ Procedure for Human Cells



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