EasySep™ Direct Human Total Lymphocyte 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
Products for Your Protocol
To see all required products for your protocol, please consult the
Protocols and Documentation.
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EasySep™ MagnetMagnet for column-free immunomagnetic separation
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D-PBS (Without Ca++ and Mg++)Dulbecco’s phosphate-buffered saline without calcium and magnesium
Overview
The EasySep™ Direct Human Total Lymphocyte Isolation Kit isolates highly purified lymphocytes directly from human whole blood by immunomagnetic negative selection. Isolated cells are untouched and available for direct use in flow cytometry, culture, DNA/RNA extraction or 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
Lymphocytes
Species
Human
Sample Source
Whole Blood
Selection Method
Negative
Application
Cell Isolation
Brand
EasySep
Area of Interest
Chimerism, HLA, Immunology
Data Figures
Figure 1. Typical EasySep™ Direct Human Total Lymphocyte Isolation Profile
Starting with human whole blood from normal healthy donors, the typical total lymphocyte (CD3-CD19+ and CD3+) content of the non-lysed final isolated fraction is 96.7 ± 1.5% (gated on CD45) or 95.8 ± 2.2% (not gated on CD45). In the example above, the total lymphocyte (CD3-CD19+ and CD3+) content of the lysed whole blood start sample and non-lysed final isolated fraction is 30.5% and 93.2% (gated on CD45), respectively, or 30.2% and 93.0% (not gated on CD45), respectively. The starting frequency of total lymphocytes in the non-lysed whole blood start sample above is 0.055% (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 1
Product Name
EasySep™ Direct Human Total Lymphocyte Isolation Kit
Catalog #
19655 Lot #
Purchase date 2020-09-28 or later Language
English Document Type
Product Information Sheet 2
Product Name
EasySep™ Direct Human Total Lymphocyte Isolation Kit
Catalog #
19655 Lot #
Purchase date before 2020-09-28 Language
English Document Type
Product Information Sheet 1
Catalog #
19655RF Lot #
Purchase date 2020-09-28 or later Language
English Document Type
Product Information Sheet 2
Catalog #
19655RF Lot #
Purchase date before 2020-09-28 Language
English Document Type
Safety Data Sheet 1
Product Name
EasySep™ Direct Human Total Lymphocyte Isolation Kit
Catalog #
19655 Lot #
All Language
English Document Type
Safety Data Sheet 2
Product Name
EasySep™ Direct Human Total Lymphocyte Isolation Kit
Catalog #
19655 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
Workflow Stages for
Total Lymphocyte Research
Workflow Stages for
HLA Analysis
Resources and Publications
Educational Materials (18)
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.
Publications (2)
Profiling of human lymphocytes reveals a specific network of protein kinases modulated by endurance training status.
Scientific reports 2020 jan
Abstract
To date, the effects of endurance exercise training on lymphocyte physiology at the kinome level are largely unknown. Therefore, the present study used a highly sensitive peptide-based kinase activity profiling approach to investigate if the basal activity of tyrosine (Tyr) and serine/threonine (Ser/Thr) kinases of human lymphocytes is affected by the aerobic endurance training status. Results revealed that the activity of various tyrosine kinases of the FGFR family and ZAP70 was increased, whereas the activity of multiple Ser/Thr kinases such as IKK$\alpha$, CaMK4, PKA$\alpha$, PKC$\alpha$+$\delta$ (among others) was decreased in lymphocytes of endurance trained athletes (ET). Moreover, functional associations between several differentially regulated kinases in ET-derived lymphocytes were demonstrated by phylogenetic mapping and network analysis. Especially, Ser/Thr kinases of the AGC-kinase (protein kinase A, G, and C) family represent exercise-sensitive key components within the lymphocytes kinase network that may mediate the long-term effects of endurance training. Furthermore, KEGG (Kyoto Encyclopedia of Genes and Genomes) and Reactome pathway analysis indicate that Ras as well as intracellular signaling by second messengers were found to be enriched in the ET individuals. Overall, our data suggest that endurance exercise training improves the adaptive immune competence by modulating the activity of multiple protein kinases in human lymphocytes.
Liquid biopsy for minimal residual disease detection in leukemia using a portable blast cell biochip.
NPJ precision oncology 2019
Abstract
Long-term management for leukemia is challenging due to the painful and invasive procedure of bone marrow (BM) biopsy. At present, non-invasive liquid (blood) biopsy is not utilized for leukemia, due to lower counts of leukemia blast cells in the blood. Here, we described a robust system for the simultaneous detection and enrichment of rare blast cells. Enrichment of blast cells was achieved from blood with a one-step microfluidic blast cell biochip (BCB) sorting system, without specific targeting of proteins by antibodies. Non-target cells encountered a differential net force as compared to stiffer blast cells and were removed. The efficiency of the BCB promotes high detection sensitivity (1 in 106 cells) even from patients with minimal residual disease. The procedure was validated using actual blast cells from patients with various types of leukemia. Outcomes were compared to current evaluation standards, such as flow cytometry, using BM aspirates. Blast cell detection efficiency was higher in 55.6{\%} of the patients using the BCB as compared to flow cytometry, despite the lower concentrations of blast cells in liquid biopsy. These studies promote early-stage detection and routine monitoring for minimal residual disease in patients.
Quality Statement:
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.
