Easily and efficiently isolate highly purified Human B Cells from HetaSep™-treated whole blood or washed leukapheresis samples by immunomagnetic negative selection, with the EasySep™ HLA B Cell Enrichment Kit: Complete Processing Kit for Whole Blood. Widely used in published research for more than 20 years, EasySep™ combines the specificity of monoclonal antibodies with the simplicity of a column-free magnetic system.
In this EasySep™ negative selection procedure, unwanted cells are labeled with antibody complexes and magnetic particles. Unwanted cells expressing the following markers are targeted for removal: CD2, CD3, CD14, CD16, CD33, CD36, CD41, CD43, CD56, CD66b, and glycophorin A. The magnetically labeled cells are then separated from the untouched desired B cells by using an EasySep™ magnet and simply pouring or pipetting the desired cells into a new tube. Following magnetic cell isolation, the desired B cells are ready for downstream applications, such as flow cytometry, culture, or DNA/RNA extraction.
Learn more about how immunomagnetic EasySep™ technology works or how to fully automate immunomagnetic cell isolation with RoboSep™ to save time and increase laboratory throughput. Explore additional products optimized for your workflow, including those for cell characterization, cryopreservation, and more.
In this EasySep™ negative selection procedure, unwanted cells are labeled with antibody complexes and magnetic particles. Unwanted cells expressing the following markers are targeted for removal: CD2, CD3, CD14, CD16, CD33, CD36, CD41, CD43, CD56, CD66b, and glycophorin A. The magnetically labeled cells are then separated from the untouched desired B cells by using an EasySep™ magnet and simply pouring or pipetting the desired cells into a new tube. Following magnetic cell isolation, the desired B cells are ready for downstream applications, such as flow cytometry, culture, or DNA/RNA extraction.
Learn more about how immunomagnetic EasySep™ technology works or how to fully automate immunomagnetic cell isolation with RoboSep™ to save time and increase laboratory throughput. Explore additional products optimized for your workflow, including those for cell characterization, cryopreservation, and more.
Data Figures
Figure 1. Typical EasySep™ HLA B Cell Enrichment Profile
Starting with HetaSep™-treated whole blood, the CD19+ cell content of the enriched fraction typically ranges from 81.5 - 99.7% (gated on CD45+ cells). Starting with 0.5 mL of HetaSep™-treated whole blood (equivalent to 5 mL of whole blood), 1.2 - 9.1 x 10e5 CD19+ cells are typically recovered.
Figure 2. FACS Data for Anti-Human CD19 Antibody, Clone HIB19, APC-Conjugated
Flow cytometry analysis of human whole blood nucleated cells processed with the EasySep™ HLA B Cell Enrichment: Complete Processing Kit for Whole Blood (Catalog #19954HLA) and labeled with Anti-Human CD19 Antibody, Clone HIB19, APC (Catalog # 60005AZ). Histograms show labeling of HetaSep™-treated whole blood cells (Start) and isolated cells (Isolated). Labeling of start cells with a mouse IgG1, kappa APC isotype control antibody is shown (open histogram).
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
RoboSep™ HLA B Cell Enrichment: Complete Processing Kit for Whole Blood with Filter Tips
Catalog #
19954HLARF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ HLA B Cell Enrichment: Complete Processing Kit for Whole Blood with Filter Tips
Catalog #
19954HLARF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ HLA B Cell Enrichment: Complete Processing Kit for Whole Blood with Filter Tips
Catalog #
19954HLARF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
RoboSep™ HLA B Cell Enrichment: Complete Processing Kit for Whole Blood with Filter Tips
Catalog #
19954HLARF
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
RoboSep™ HLA B Cell Enrichment: Complete Processing Kit for Whole Blood with Filter Tips
Catalog #
19954HLARF
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 B Cell Research
Workflow Stages for HLA Analysis
Resources and Publications
Educational Materials (8)
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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