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EasySep™ Direct Human Monocyte Isolation Kit

Immunomagnetic negative selection from whole blood kit

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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™ Direct Human Monocyte Isolation Kit

Immunomagnetic negative selection from whole blood kit

From: 567 USD
Catalog #
19669_C
Immunomagnetic negative selection from whole blood kit
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Product Advantages


  • 99.9% RBC depletion without the need for density gradient centrifugation, sedimentation, or lysis

  • Up to 92% purity of isolated cells

  • Fast, easy-to-use and column-free

  • Isolated cells are untouched

What's Included

  • EasySep™ Direct Human Monocyte Isolation Kit (Catalog #19669)
    • EasySep™ Direct Human Monocyte Isolation Cocktail, 2 x 2.5 mL
    • EasySep™ Direct RapidSpheres™, 4 x 2.5 mL
  • RoboSep™ Human Monocyte Isolation Kit with Filter Tips (Catalog #19669RF)
    • EasySep™ Direct Human Monocyte Isolation Cocktail, 2 x 2.5 mL
    • EasySep™ Direct RapidSpheres™, 4 x 2.5 mL
    • RoboSep™ Buffer (Catalog #20104)
    • RoboSep™ Filter Tips (Catalog #20125) x 2
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
Products for Your Protocol
To see all required products for your protocol, please consult the Protocols and Documentation.
  1. EasySep™ Magnet
    EasySep™ Magnet

    Magnet for column-free immunomagnetic separation

  2. D-PBS (Without Ca++ and Mg++)
    D-PBS (Without Ca++ and Mg++)

    Dulbecco’s phosphate-buffered saline without calcium and magnesium

Overview

The EasySep™ Direct Human Monocyte Isolation Kit isolates functional, highly purified CD14+ monocytes directly from human whole blood by immunomagnetic negative selection. No lysis, density gradient centrifugation or other processing steps are required and isolated cells are immediately available for flow cytometry, functional assays, culture and other downstream applications.
Magnet Compatibility
• EasySep™ Magnet (Catalog #18000)
• “The Big Easy” EasySep™ Magnet (Catalog #18001)
• Easy 50 EasySep™ Magnet (Catalog #18002)
• EasyEights™ EasySep™ Magnet (Catalog #18103)
• RoboSep™-S (Catalog #21000)
Subtype
Cell Isolation Kits
Cell Type
Monocytes, Myeloid Cells
Species
Human
Sample Source
Whole Blood
Selection Method
Negative
Application
Cell Isolation
Brand
EasySep, RoboSep
Area of Interest
Immunology

Data Figures

Monocyte separation using EasySep™ Direct Human Monocyte Isolation Kit

Figure 1. Typical EasySep™ Direct Human Monocyte Isolation Profile

Starting with human whole blood from normal healthy donors, the typical monocyte (CD14+) content of the non-lysed final Isolated fraction is 82.2 ± 8.4% (gated on CD45) or 79.0 ± 10.1% (not gated on CD45). In the example above, the monocyte (CD14+) content of the lysed whole blood start sample and the non-lysed final isolated fraction is 6.5% and 88.3% (gated on CD45), respectively, or 6.4% and 85.7% (not gated on CD45), respectively. The starting frequency of monocytes in the non-lysed whole blood start sample above is approximately 0.007% (data not shown).

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
Document Type
Product Information Sheet
Product Name
EasySep™ Direct Human Monocyte Isolation Kit
Catalog #
19669
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
EasySep™ Direct Human Monocyte Isolation Kit for RoboSep™
Catalog #
19669RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Direct Human Monocyte Isolation Kit
Catalog #
19669
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Direct Human Monocyte Isolation Kit
Catalog #
19669
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Direct Human Monocyte Isolation Kit for RoboSep™
Catalog #
19669RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Direct Human Monocyte Isolation Kit for RoboSep™
Catalog #
19669RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
EasySep™ Direct Human Monocyte Isolation Kit for RoboSep™
Catalog #
19669RF
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.

Research Area
Workflow Stages

Resources and Publications

Educational Materials (11)

EasySep™ Direct: Cell Isolation from Whole Blood in 20 Minutes
Brochure
EasySep™ Direct: Cell Isolation from Whole Blood in 20 Minutes
Tools to Streamline your Macrophage Research
Brochure
Tools to Streamline your Macrophage Research
EasySep™ Cell Separation Technology
Brochure
EasySep™ Cell Separation Technology
Frequencies of Human Cell Types in Blood-Related Sources
Wallchart
Frequencies of Human Cell Types in Blood-Related Sources
Antigen Processing and Presentation
Wallchart
Antigen Processing and Presentation
Human Immune Cytokines
Wallchart
Human Immune Cytokines
How to Isolate Cells Directly from Whole Blood Using the EasySep™ Purple/Silver Magnets
4:30
Video
How to Isolate Cells Directly from Whole Blood Using the EasySep™ Purple/Silver Magnets
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology
1:13
Video
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology
Cell Isolation Directly from Whole Blood without RBC Lysis or Centrifugation: EasySep™ Direct
1:44
Video
Cell Isolation Directly from Whole Blood without RBC Lysis or Centrifugation: EasySep™ Direct
How to Isolate Cells Directly from Whole Blood Using the Easy 50 EasySep™ Magnet: 30-Minute Protocol
5:17
Video
How to Isolate Cells Directly from Whole Blood Using the Easy 50 EasySep™ Magnet: 30-Minute Protoc...
How to Isolate Cells Directly from Whole Blood - EasySep™ Direct Protocol, EasyEights Magnet
6:13
Video
How to Isolate Cells Directly from Whole Blood - EasySep™ Direct Protocol, EasyEights Magnet

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)

S100-alarmin-induced innate immune programming protects newborn infants from sepsis. T. Ulas et al. Nature immunology 2017 MAY

Abstract

The high risk of neonatal death from sepsis is thought to result from impaired responses by innate immune cells; however, the clinical observation of hyperinflammatory courses of neonatal sepsis contradicts this concept. Using transcriptomic, epigenetic and immunological approaches, we demonstrated that high amounts of the perinatal alarmins S100A8 and S100A9 specifically altered MyD88-dependent proinflammatory gene programs. S100 programming prevented hyperinflammatory responses without impairing pathogen defense. TRIF-adaptor-dependent regulatory genes remained unaffected by perinatal S100 programming and responded strongly to lipopolysaccharide, but were barely expressed. Steady-state expression of TRIF-dependent genes increased only gradually during the first year of life in human neonates, shifting immune regulation toward the adult phenotype. Disruption of this critical sequence of transient alarmin programming and subsequent reprogramming of regulatory pathways increased the risk of hyperinflammation and sepsis. Collectively these data suggest that neonates are characterized by a selective, transient microbial unresponsiveness that prevents harmful hyperinflammation in the delicate neonate while allowing for sufficient immunological protection.
View publication
Germline-encoded neutralization of a Staphylococcus aureus virulence factor by the human antibody repertoire. Yeung YA et al. Nature communications 2016 NOV

Abstract

Staphylococcus aureus is both an important pathogen and a human commensal. To explore this ambivalent relationship between host and microbe, we analysed the memory humoral response against IsdB, a protein involved in iron acquisition, in four healthy donors. Here we show that in all donors a heavily biased use of two immunoglobulin heavy chain germlines generated high affinity (pM) antibodies that neutralize the two IsdB NEAT domains, IGHV4-39 for NEAT1 and IGHV1-69 for NEAT2. In contrast to the typical antibody/antigen interactions, the binding is primarily driven by the germline-encoded hydrophobic CDRH-2 motifs of IGHV1-69 and IGHV4-39, with a binding mechanism nearly identical for each antibody derived from different donors. Our results suggest that IGHV1-69 and IGHV4-39, while part of the adaptive immune system, may have evolved under selection pressure to encode a binding motif innately capable of recognizing and neutralizing a structurally conserved protein domain involved in pathogen iron acquisition.
View publication
View All Publications

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