EasySep™ Human Memory CD8+ T Cell Enrichment Kit

Immunomagnetic negative selection kit
Catalog #
19159_C
Immunomagnetic negative selection kit
From: 685 USD
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Required Products
  1. EasySep™ Magnet
    EasySep™ Magnet

    Magnet for column-free immunomagnetic separation

  2. EasySep™ Buffer
    EasySep™ Buffer

    Cell separation buffer

Overview

The EasySep™ Human Memory CD8+ T Cell Enrichment Kit is designed to isolate memory CD8+ T cells from peripheral blood mononuclear cells by negative selection. Unwanted cells are targeted for removal with antibody complexes recognizing CD4, CD14, CD16, CD19, CD20, CD34, CD36, CD45RA, CD56, CD61, CD66b, CD123, TCRγ/δ, glycophorin A 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.
Advantages
• Fast, easy-to-use and column-free
• Up to 92% purity
• Untouched, viable cells
Components
  • EasySep™ Human Memory CD8+ T Cell Enrichment Kit (Catalog #19159)
    • EasySep™ Memory CD8+ T Cell Enrichment Cocktail, 1 mL
    • EasySep™ Magnetic Particles, 3 x 1 mL
  • RoboSep™ Human Memory CD8+ T Cell Enrichment Kit (Catalog #19159RF)
    • EasySep™ Memory CD8+ T Cell Enrichment Cocktail, 1 mL
    • EasySep™ Magnetic Particles, 3 x 1 mL
    • RoboSep™ Buffer (Catalog #20104)
    • RoboSep™ Filter Tips (Catalog #20125)
Magnet Compatibility
• EasySep™ Magnet (Catalog #18000)
• “The Big Easy” EasySep™ Magnet (Catalog #18001)
• RoboSep™-S (Catalog #21000)
Subtype
Cell Isolation Kits
Cell Type
T Cells, T Cells, CD8+
Species
Human
Sample Source
Leukapheresis, PBMC
Selection Method
Negative
Application
Cell Isolation
Brand
EasySep, RoboSep
Area of Interest
Immunology

Related Products

Labeling Antibodies

Scientific Resources

Product Documentation

Document Type Product Name Catalog # Lot # Language
Document Type
Product Information Sheet
Product Name
EasySep™ Human Memory CD8+ T Cell Enrichment Kit
Catalog #
19159
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
RoboSep™ Human Memory CD8+ T Cell Enrichment Kit
Catalog #
19159RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Human Memory CD8+ T Cell Enrichment Kit
Catalog #
19159
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Human Memory CD8+ T Cell Enrichment Kit
Catalog #
19159
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ Human Memory CD8+ T Cell Enrichment Kit
Catalog #
19159RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ Human Memory CD8+ T Cell Enrichment Kit
Catalog #
19159RF
Lot #
All
Language
English

Educational Materials(7)

Brochure
Human T Cell Research Product Workflow
Brochure
Tools For Your Immunology Research
Brochure
EasySep™ Cell Separation Technology
Wallchart
Human Immune Cytokines
Video
1:13
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology
Video
1:57
How EasySep™ Magnetic Cell Separation Technology Works: Fast and Easy Cell Isolation
Video
1:37
How to Isolate PBMCs from Whole Blood Using Density Gradient Centrifugation (Ficoll™ or Lymphoprep™)

Frequently Asked Question

Can EasySep™ be used for either positive or negative selection?

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).

How does the separation work?

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.

Which columns do I use?

The EasySep™ procedure is column-free. That's right - no columns!

How can I analyze the purity of my enriched sample?

The Product Information Sheet provided with each EasySep™ kit contains detailed staining information.

Can EasySep™ separations be automated?

Yes. RoboSep™, the fully automated cell separator, automates all EasySep™ labeling and cell separation steps.

Can EasySep™ be used to isolate rare cells?

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.

Are the EasySep™ magnetic particles FACS-compatible?

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.

Can the EasySep™ magnetic particles be removed after enrichment?

No, but due to the small size of these particles, they will not interfere with downstream applications.

Can I alter the separation time in the magnet?

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.

For positive selection, can I perform more than 3 separations to increase purity?

Yes, the purity of targeted cells will increase with additional rounds of separations; however, cell recovery will decrease.

How does the binding of the EasySep™ magnetic particle affect the cells? is the function of positively selected cells altered by the bound particles?

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.
Read More

Product Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Data and Publications

Data

Typical EasySep™ Human Memory CD8+ T Cell Enrichment Profile

Figure 1. Typical EasySep™ Human Memory CD8+ T Cell Enrichment Profile

Starting with mononuclear cells, the memory CD8+ T cell (CD8+CD45RA-CD45RO+) content of the enriched fraction typically ranges from 72 - 92%. In the example above, the purities of the start and enriched fractions are 4.9% and 88.8%, respectively.

Publications (1)

Nature communications 2018 OCT Human breast tumor-infiltrating CD8+ T cells retain polyfunctionality despite PD-1 expression. C. A. Egelston et al.

Abstract

Functional CD8+ T cells in human tumors play a clear role in clinical prognosis and response to immunotherapeutic interventions. PD-1 expression in T cells involved in chronic infections and tumors such as melanoma often correlates with a state of T-cell exhaustion. Here we interrogate CD8+ tumor-infiltrating lymphocytes (TILs) from human breast and melanoma tumors to explore their functional state. Despite expression of exhaustion hallmarks, such as PD-1 expression, human breast tumor CD8+ TILs retain robust capacity for production of effector cytokines and degranulation capacity. In contrast, melanoma CD8+ TILs display dramatic reduction of cytokine production and degranulation capacity. We show that CD8+ TILs from human breast tumors can potently kill cancer cells via bi-specific antibodies. Our data demonstrate that CD8+ TILs in human breast tumors retain polyfunctionality, despite PD-1 expression, and suggest that they may be harnessed for effective immunotherapies.
View All Publications

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