EasySep™ Human CD3 Positive Selection Kit II

Immunomagnetic positive selection kit

New look, same high quality and support! You may notice that your instrument or reagent packaging looks slightly different from images displayed on the website, or from previous orders. We are updating our look but rest assured, the products themselves and how you should use them have not changed. Learn more

EasySep™ Human CD3 Positive Selection Kit II

Immunomagnetic positive selection kit

From: 771 USD
Catalog #
17851_C
Immunomagnetic positive selection kit

Product Advantages


  • Fast and easy-to-use

  • Up to 99% purity

  • No columns required

What's Included

  • EasySep™ Human CD3 Positive Selection Kit II (Catalog #17851)
    • EasySep™ Human CD3 Positive Selection Cocktail II, 1 mL
    • EasySep™ Dextran RapidSpheres™ 50100, 1 mL
  • EasySep™ Human CD3 Positive Selection Kit II (Catalog #100-0692)
    • EasySep™ Human CD3 Positive Selection Cocktail II, 1 x 10 mL
    • EasySep™ Dextran RapidSpheres™ 50103, 2 x 1 mL
  • RoboSep™ Human CD3 Positive Selection Kit II (Catalog #17851RF)
    • EasySep™ Human CD3 Positive Selection Cocktail II, 1 mL
    • EasySep™ Dextran RapidSpheres™ 50100, 1 mL
    • RoboSep™ Buffer (Catalog #20104)
    • RoboSep™ Filter Tips (Catalog #20125)
New look, same high quality and support! You may notice that your instrument or reagent packaging looks slightly different from images displayed on the website, or from previous orders. We are updating our look but rest assured, the products themselves and how you should use them have not changed. Learn more

Overview

The EasySep™ Human CD3 Positive Selection Kit II is designed to isolate CD3+ cells from fresh or previously frozen peripheral blood mononuclear cells by positive selection. Desired cells are targeted with Tetrameric Antibody Complexes recognizing CD3 and dextran-coated magnetic particles. The cocktail also contains an antibody to human Fc receptor to minimize nonspecific binding. Labeled cells are separated using an EasySep™ magnet without the use of columns. Cells of interest remain in the tube while unwanted cells are poured off. The CD3 antigen is expressed on all T cells and CD56+ NKT cells.

This product replaces the EasySep™ Human CD3 Positive Selection Kit (Catalog #18051) for even faster cell isolations.
Magnet Compatibility
• EasySep™ Magnet (Catalog #18000)
• “The Big Easy” EasySep™ Magnet (Catalog #18001)
• EasyPlate™ EasySep™ Magnet (Catalog #18102)
• EasyEights™ EasySep™ Magnet (Catalog #18103)
• Easy 50 EasySep™ Magnet (Catalog #18002)
• RoboSep™-S (Catalog #21000)
• Easy 250 EasySep™ Magnet (Catalog #100-0821)
Subtype
Cell Isolation Kits
Cell Type
T Cells
Species
Human
Sample Source
PBMC
Selection Method
Positive
Application
Cell Isolation
Brand
EasySep, RoboSep
Area of Interest
Cell Therapy Research, Chimerism, Immunology

Data Figures

Typical FACS Results with EasySep™ Human CD3 Positive Selection Kit II

Figure 1: Typical FACS Results with EasySep™ Human CD3 Positive Selection Kit II

Starting with a single cell suspension of human PBMCs, the CD3+ cell content of the isolated fraction is typically 99.2 ± 0.2% (mean ± SD using the purple EasySep™ Magnet).

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

Document Type
Product Name
Catalog #
Lot #
Language
Catalog #
17851
Lot #
All
Language
English
Catalog #
17851RF
Lot #
All
Language
English
Catalog #
100-0692
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
17851
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
17851
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
17851RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
17851RF
Lot #
All
Language
English
Document Type
Safety Data Sheet
Catalog #
100-0692
Lot #
All
Language
English

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.

Resources and Publications

Frequently Asked Questions

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.

Publications (2)

BiHC, a T-Cell-Engaging Bispecific Recombinant Antibody, Has Potent Cytotoxic Activity Against Her2 Tumor Cells. Xing J et al. Translational oncology 2017 OCT

Abstract

Among different cancer immunotherapy approaches, bispecific antibodies (BsAbs) are of great interest due to their ability to recruit immune cells to kill tumor cells directly. Various BsAbs against Her2 tumor cells have been proposed with potent cytotoxic activities. However, most of these formats require extensive processing to obtain heterodimeric bispecific antibodies. In this study, we describe a bispecific antibody, BiHC (bispecific Her2-CD3 antibody), constructed with a single-domain anti-Her2 and a single-chain Fv (variable fragment) of anti-CD3 in an IgG-like format. In contrast to most IgG-like BsAbs, the two arms in BiHC have different molecular weights, making it easier to separate hetero- or homodimers. BiHC can be expressed in Escherichia coli and purified via Protein A affinity chromatography. The purified BiHC can recruit T cells and induce specific cytotoxicity of Her2-expressing tumor cells in vitro. The BiHC can also efficiently inhibit the tumor growth in vivo. Thus, BiHC is a promising candidate for the treatment of Her2-positive cancers.
An optimized protocol for adenosine triphosphate quantification in T lymphocytes of lymphopenic patients. Girardot T et al. Journal of immunological methods 2016 OCT

Abstract

In several clinical contexts, the measurement of ATP concentration in T lymphocytes has been proposed as a biomarker of immune status, predictive of secondary infections. However, the use of such biomarker in lymphopenic patients requires some adaptations in the ATP dosage protocol. We used blood from healthy volunteers to determine the optimal experimental settings. We investigated technical aspects such as the type of anticoagulant for blood sampling, the effect of freeze and thaw cycles, the reagent and sample mixing sequence, and the optimal dilution buffer. We also shortened the incubation time to 8h, and even showed that a 30min incubation may be sufficient. To evaluate the ATP rise upon lymphocyte activation, the optimal dose of stimulant was defined to be 4μg/mL of phytohaemagglutinin. Lastly, we determined that the number of T cells needed for this measurement was as low as 50,000, which is compatible with the existing lymphopenia in clinical settings. This optimized protocol appears ready to be assessed in lymphopenic patients to further investigate the interconnection between T lymphocyte metabolism and impaired phenotype and functions.

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