EasySep™ Human Pan-B Cell Enrichment Kit

Immunomagnetic negative selection cell isolation kit

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EasySep™ Human Pan-B Cell Enrichment Kit

Immunomagnetic negative selection cell isolation kit

From: 861 USD
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Immunomagnetic negative selection cell isolation kit
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Product Advantages


  • Fast, easy-to-use and column-free

  • Up to 99% purity

  • Isolated cells are untouched

What's Included

  • EasySep™ Human Pan-B Cell Enrichment Kit (Catalog #19554)
    • EasySep™ Human Pan-B Cell Enrichment Cocktail, 1 mL
    • EasySep™ Magnetic Particles, 2 x 1 mL
  • RoboSep™ Human Pan-B Cell Enrichment Kit (Catalog #19554RF)
    • EasySep™ Human Pan-B Cell Enrichment Cocktail, 1 mL
    • EasySep™ Magnetic Particles, 2 x 1 mL
    • RoboSep™ Buffer (Catalog #20104)
    • RoboSep™ Filter Tips (Catalog #20125)

Overview

The EasySep™ Human Pan-B Cell Enrichment Kit is designed to isolate B cells including plasma cells from normal fresh or previously frozen peripheral blood mononuclear cells by negative selection. Undesired cells are targeted for removal with Tetrameric Antibody Complexes recognizing CD2, CD3, CD14, CD16, CD36, CD42b, CD56, CD66b, CD123, glycophorin A and dextran-coated magnetic particles. The labeled cells are separated using an EasySep™ magnet without the use of columns. Desired cells are poured off into a new tube.
Magnet Compatibility
• EasySep™ Magnet (Catalog #18000)
• “The Big Easy” EasySep™ Magnet (Catalog #18001)
• Easy 50 EasySep™ Magnet (Catalog #18002)
• RoboSep™-S (Catalog #21000)
Subtype
Cell Isolation Kits
Cell Type
B Cells, Plasma
Species
Human
Sample Source
Leukapheresis, PBMC
Selection Method
Negative
Application
Cell Isolation
Brand
EasySep, RoboSep
Area of Interest
Immunology

Data Figures

FACS Profile Results With EasySep™ Human Pan-B Cell Enrichment Kit

Figure 1. FACS Profile Results With EasySep™ Human Pan-B Cell Enrichment Kit

Starting with nucleated cells, the pan-B cell [Lineage (CD4, CD8, CD14, CD16, CD56) negative, CD19+ and CD19-CD43+] content of the enriched fraction typically ranges from 90 - 99%.

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
Catalog #
19554RF
Lot #
All
Language
English
Catalog #
19554
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
19554RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
19554RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Catalog #
19554RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
19554
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
19554
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.

Resources and Publications

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.

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.

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.

Which cell separation kits are compatible with the "Easy 50" EASYSEP™ magnet?

At present, the "Easy 50" EasySep™ magnet is only compatible with EasySep™ kits for human cell separation.

T Cells: 19051 (T Cells), 19052 (CD4 T cells), 19157 (Memory CD4 T Cells), 19053 (CD8 T Cells), 19159 (Memory CD8 T Cells - please contact Tech Support)

B Cells: 19054 (B Cells), 19254 (Naïve B cells)

Other Cell Types: 19055 (NK Cells), 19058 (Monocytes without CD16 depletion), 19059 (Monocytes), 19062 (Plasmacytoid DCs), 19251 (pan-DCs)

For HLA Analysis: 19951HLA (T Cells from whole blood), 19954HLA (B Cells from whole blood), 19961HLA (Total lymphocytes from whole blood)

Publications (2)

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.
HIF-1α regulates the interaction of chronic lymphocytic leukemia cells with the tumor microenvironment. Valsecchi R et al. Blood 2016 APR

Abstract

Hypoxia-inducible transcription factors (HIFs) regulate a wide array of adaptive responses to hypoxia and are often activated in solid tumors and hematologic malignancies due to intratumoral hypoxia and emerging new layers of regulation. We found that in chronic lymphocytic leukemia (CLL), HIF-1α is a novel regulator of the interaction of CLL cells with protective leukemia microenvironments and, in turn, is regulated by this interaction in a positive feedback loop that promotes leukemia survival and propagation. Through unbiased microarray analysis, we found that in CLL cells, HIF-1α regulates the expression of important chemokine receptors and cell adhesion molecules that control the interaction of leukemic cells with bone marrow and spleen microenvironments. Inactivation of HIF-1α impairs chemotaxis and cell adhesion to stroma, reduces bone marrow and spleen colonization in xenograft and allograft CLL mouse models, and prolongs survival in mice. Of interest, we found that in CLL cells, HIF-1α is transcriptionally regulated after coculture with stromal cells. Furthermore, HIF-1α messenger RNA levels vary significantly within CLL patients and correlate with the expression of HIF-1α target genes, including CXCR4, thus further emphasizing the relevance of HIF-1α expression to CLL pathogenesis.
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