EasySep® Human Memory B Cell Isolation Kit

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Immunomagnetic Positive Selection of Human Memory B Cells

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  • Box top label for Human Memory B Cell Isolation Kit
EasySep® box
The EasySep® Human Memory B Cell Isolation Kit is a two-step method designed to isolate CD27+ B cells from fresh or previously frozen peripheral blood nucleated cells. B cells are first pre-enriched by negative selection using EasySep® Human Memory B Cell Enrichment Cocktail. Unwanted cells are targeted for removal with Tetrameric Antibody Complexes (TAC) recognizing CD2, CD3, CD14, CD16, CD36, CD43, CD56, CD66b, glycophorinA and dextran-coated magnetic particles. The labeled cells are separated using an EasySep® magnet without the use of columns. Desired cells are poured off into a new tube. Following pre-enrichment, CD27 positive B cells are selected using EasySep® Human CD27 Positive Selection Cocktail. Cells are targeted with TAC recognizing CD27 and dextran-coated magnetic particles. Labeled cells are separated using an EasySep® magnet and remain in the tube while the unwanted cells are poured off. This negative cell fraction can be used to obtain naïve B cells. The selection steps can be fully automated using RoboSep®.
Product Name Description Catalog # Size Price Quantity
EasySep® Human Memory B Cell Isolation Kit Immunomagnetic positive selection of human memory B cells 18164 For labeling up to 2 x 10e9 cells 788.00 USD      
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RoboSep® Human Memory B Cell Isolation Kit EasySep® kit with RoboSep® buffer and RoboSep® filter tips 18164RF For labeling up to 2 x 10e9 cells 820.00 USD      
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Order Offline:

Call us: +1.604.877.0713  
Toll Free: 1.800.667.0322  
Email us:
 
Fax us: +1.604.877.0704
Toll Free Fax: 1.800.567.2899

Recommended for:
Isolation of memory B cells (CD27+) from fresh or previously frozen peripheral blood nucleated cells.
Components:
• EasySep® Human B Cell Enrichment Cocktail, 2 x 1.0 ml
• EasySep® Magnetic Particles, 5 x 1.0 ml
• EasySep® Human CD27 Positive Selection Cocktail, 2 x 1.0 ml
• EasySep® Magnetic Particles, 1.0 ml
 
Intended Use Statement: For Research Use Only. Not for Therapeutic or Diagnostic Use.
Equipment Required:
• EasySep® Magnet (Catalog #18000), or
• “The Big Easy” EasySep® Magnet (Catalog #18001), or
• RoboSep® (Catalog #20000)
Product Type: Cell isolation products
Area of Interest: Immunology
Cell Isolation Product Type: Reagents
Cell Source: PBMC
Cell Type: B cells
Popular Product Line: EasySep
Selection Method: Positive
Species: Human

Procedures and instruction manuals:

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.

Product Name

Description

Catalog #
EasySep® Magnet Immunomagnetic Column-Free Magnet 18000
The Big Easy EasySep® Magnet Immunomagnetic Column-Free Magnet 18001
EasySep® Human B Cell Enrichment Kit Immunomagnetic Negative Selection Kit 19054
EasySep® Human Naive B Cell Enrichment Kit Immunomagnetic Negative Selection Kit 19254
RoboSep™ RoboSep™ - The Fully Automated Cell Separator 20000

Typical EasySep® Human Memory B Cell Selection Profile


Typical EasySep® Human Memory B Cell Selection Profile
Starting with nucleated cells, the CD19+CD27+ cell content of the enriched fraction typically ranges from 85 - 95%.


Typical EasySep® Procedure for Human Cells


Typical EasySep® Procedure for Human Cells



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