EasySep™ Human CD14 Positive Selection Kit II

Immunomagnetic positive selection cell isolation 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 CD14 Positive Selection Kit II

Immunomagnetic positive selection cell isolation kit

From: 790 USD
Catalog #
17858_C
Immunomagnetic positive selection cell isolation kit

Product Advantages

• Fast and easy-to-use
• Up to 97% purity
• No columns required

What's Included

  • EasySep™ Human CD14 Positive Selection Kit II (Catalog #17858)
    • EasySep™ Human CD14 Positive Selection Cocktail II, 1 mL
    • EasySep™ Dextran RapidSpheres™ 50100, 1 mL
  • EasySep™ Human CD14 Positive Selection Kit II (Catalog #100-0694)
    • EasySep™ Human CD14 Positive Selection Cocktail II, 1 x 10 mL
    • EasySep™ Dextran RapidSpheres™ 50103, 2 x 1 mL
  • RoboSep™ Human CD14 Positive Selection Kit II (Catalog #17858RF)
    • EasySep™ Human CD14 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 CD14 Positive Selection Kit II is designed to isolate highly purified CD14+ cells from fresh or previously frozen human peripheral blood mononuclear cells (PBMCs). Desired cells are targeted with Tetrameric Antibody Complexes recognizing CD14 and dextran-coated magnetic particles. 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 CD14 antigen is strongly expressed on monocytes and macrophages and weakly on granulocytes. It is also expressed on most tissue macrophages.

This product replaces the EasySep™ Human CD14 Positive Selection Kit (Catalog #18058), for even faster cell isolations and is compatible with all EasySep™ magnets.
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
Monocytes, Myeloid Cells
Species
Human
Sample Source
PBMC
Selection Method
Positive
Application
Cell Isolation
Brand
EasySep, RoboSep
Area of Interest
Chimerism, Immunology

Scientific Resources

Product 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 #
17858
Lot #
1000087738 or lower
Language
English
Catalog #
17858
Lot #
1000087739 or higher
Language
English
Catalog #
17858RF
Lot #
1000087738 or lower
Language
English
Catalog #
17858RF
Lot #
1000087739 or higher
Language
English
Document Type
Safety Data Sheet 1
Catalog #
17858
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
17858
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
17858RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
17858RF
Lot #
All
Language
English
Document Type
Safety Data Sheet
Catalog #
100-0694
Lot #
All
Language
English

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.

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

Figure 1. Typical EasySep™ Human CD14 Positive Selection II Isolation Profile

Starting with a single cell suspension of human PBMCs, the CD14+ cell content of the isolated fraction is typically 95.3 ± 4.5% (mean ± SD using the purple EasySep™ Magnet).

Publications (20)

Illuminating Host-Mycobacterial Interactions with Genome-wide CRISPR Knockout and CRISPRi Screens. Y. Lai et al. Cell systems 2020 sep

Abstract

Existing antibiotics are inadequate to defeat tuberculosis (TB), a leading cause of death worldwide. We sought potential targets for host-directed therapies (HDTs) by investigating the host immune response to mycobacterial infection. We used high-throughput CRISPR knockout and CRISPR interference (CRISPRi) screens to identify perturbations that improve the survival of human phagocytic cells infected with Mycobacterium bovis BCG (Bacillus Calmette-Gu{\'{e}}rin), as a proxy for Mycobacterium tuberculosis (Mtb). Many of these perturbations constrained the growth of intracellular mycobacteria. We identified over 100 genes associated with diverse biological pathways as potential HDT targets. We validated key components of the type I interferon and aryl hydrocarbon receptor signaling pathways that respond to the small-molecule inhibitors cerdulatinib and CH223191, respectively; these inhibitors enhanced human macrophage survival and limited the intracellular growth of Mtb. Thus, high-throughput functional genomic screens, by elucidating highly complex host-pathogen interactions, can serve to identify HDTs to potentially improve TB treatment.
Detecting Tumor Antigen-Specific T Cells via Interaction-Dependent Fucosyl-Biotinylation. Z. Liu et al. Cell 2020 nov

Abstract

Re-activation and clonal expansion of tumor-specific antigen (TSA)-reactive T cells are critical to the success of checkpoint blockade and adoptive transfer of tumor-infiltrating lymphocyte (TIL)-based therapies. There are no reliable markers to specifically identify the repertoire of TSA-reactive T cells due to their heterogeneous composition. We introduce FucoID as a general platform to detect endogenous antigen-specific T cells for studying their biology. Through this interaction-dependent labeling approach, intratumoral TSA-reactive CD4+, CD8+ T cells, and TSA-suppressive CD4+ T cells can be detected and separated from bystander T cells based on their cell-surface enzymatic fucosyl-biotinylation. Compared to bystander TILs, TSA-reactive TILs possess a distinct T cell receptor (TCR) repertoire and unique gene features. Although exhibiting a dysfunctional phenotype, TSA-reactive CD8+ TILs possess substantial capabilities of proliferation and tumor-specific killing. Featuring genetic manipulation-free procedures and a quick turnover cycle, FucoID should have the potential of accelerating the pace of personalized cancer treatment.
Impact of mRNA chemistry and manufacturing process on innate immune activation. J. Nelson et al. Science advances 2020 jun

Abstract

Messenger RNA (mRNA) represents an attractive therapeutic modality for potentially a wide range of clinical indications but requires uridine chemistry modification and/or tuning of the production process to prevent activation of cellular innate immune sensors and a concomitant reduction in protein expression. To decipher the relative contributions of these factors on immune activation, here, we compared, in multiple cell and in vivo models, mRNA that encodes human erythropoietin incorporating either canonical uridine or N1-methyl-pseudouridine (1m$\Psi$), synthesized by either a standard process shown to have double-stranded RNA (dsRNA) impurities or a modified process that yields a highly purified mRNA preparation. Our data demonstrate that the lowest stimulation of immune endpoints was with 1m$\Psi$ made by the modified process, while mRNA containing canonical uridine was immunostimulatory regardless of process. These findings confirm that uridine modification and the reduction of dsRNA impurities are both necessary and sufficient at controlling the immune-activating profile of therapeutic mRNA.

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