EasySep™ Direct Human Monocyte Isolation Kit
Immunomagnetic negative selection from whole blood kit
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EasySep™ Direct Human Monocyte Isolation Kit -
Label for EasySep™ Direct Human Monocyte Isolation Kit
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Required Products
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Magnet for column-free immunomagnetic separation -
Dulbecco’s phosphate-buffered saline without calcium and magnesium -
Compatible antibodies for purity assessment of isolated cells
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.
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
• Up to 92% purity of isolated cells
• Fast, easy-to-use and column-free
• Isolated cells are untouched
Components:
- 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
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
Related Products
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EasySep™ Human Monocyte Isolation Kit
12.5-Minute cell isolation kit using immunomagnetic negative selection -
Human Recombinant GM-CSF (E. coli-expressed)
Granulocyte-macrophage colony-stimulating factor -
Primary human cells, fresh -
ImmunoCult™ Dendritic Cell Culture Kit
For differentiation of human monocytes into dendritic cells
Labeling Antibodies
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Anti-Human CD14 Antibody, Clone M5E2
Mouse monoclonal IgG2a antibody against human, rhesus, cynomolgus CD14
Scientific Resources
Product Documentation
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(12)
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.
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.
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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.
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Workflow Stages for
Workflow Stages
Data and Publications
Data
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).
Publications
(2)
Nature immunology 2017 MAY
S100-alarmin-induced innate immune programming protects newborn infants from sepsis.
Abstract
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.
Nature communications 2016 NOV
Germline-encoded neutralization of a Staphylococcus aureus virulence factor by the human antibody repertoire.
Abstract
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.
PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.