EasySep™ Mouse B Cell Isolation Kit

15-Minute cell isolation kit using immunomagnetic negative selection
Catalog #
19854_C
15-Minute cell isolation kit using immunomagnetic negative selection
From: 887 EUR
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Required Products
  1. EasyEights™ EasySep™ Magnet
    EasyEights™ EasySep™ Magnet

    Magnet for column-free immunomagnetic separation

  2. EasySep™ Buffer
    EasySep™ Buffer

    Cell separation buffer

Overview

The EasySep™ Mouse B Cell Isolation Kit is designed to isolate B cells from single-cell suspensions of splenocytes or other tissues by negative selection. Unwanted cells are targeted for removal with biotinylated antibodies directed against non-B cells and streptavidin-coated magnetic particles (RapidSpheres™ ). Labeled cells are separated using and EasySep™ magnet without the use of columns. Desired cells are poured off into a new tube.

For isolation of B cells expressing CD11b or CD43, we recommend using the EasySep™ Mouse Pan-B Cell Isolation kit (Catalog #19844).

This product replaces the EasySep™ Mouse B Cell Enrichment Kit (Catalog #19754) for even faster cell isolations.
Advantages
• Fast and easy-to-use
• Up to 95% purity
• No columns required
• Untouched, viable cells
Components
  • EasySep™ Mouse B Cell Isolation Kit (Catalog #19854)
    • EasySep™ Mouse B Cell Isolation Cocktail, 0.5 mL
    • EasySep™ Streptavidin RapidSpheres™ 50001, 1 mL
    • Normal Rat Serum, 2 mL
  • RoboSep™ Mouse B Cell Isolation Kit (Catalog #19854RF)
    • EasySep™ Mouse B Cell Isolation Cocktail, 0.5 mL
    • EasySep™ Streptavidin RapidSpheres™ 50001, 1 mL
    • Normal Rat Serum, 2 mL
    • RoboSep™ Buffer (Catalog #20104)
    • RoboSep™ Filter Tips (Catalog #20125)
Magnet Compatibility
• EasySep™ Magnet (Catalog #18000)
• “The Big Easy” EasySep™ Magnet (Catalog #18001)
• EasyPlate™ EasySep™ Magnet (Catalog 18102)
• EasyEights™ EasySep™ Magnet (Catalog #18103)
• RoboSep™-S (Catalog #21000)
Subtype
Cell Isolation Kits
Cell Type
B Cells
Species
Mouse
Sample Source
Other, Spleen
Selection Method
Negative
Application
Cell Isolation
Brand
EasySep, RoboSep
Area of Interest
Immunology

Related Products

Labeling Antibodies

Scientific Resources

Product Documentation

Document Type Product Name Catalog # Lot # Language
Document Type
Product Information Sheet
Product Name
EasySep™ Mouse B Cell Isolation Kit
Catalog #
19854
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
RoboSep™ Mouse B Cell Isolation Kit
Catalog #
19854RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Mouse B Cell Isolation Kit
Catalog #
19854
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Mouse B Cell Isolation Kit
Catalog #
19854
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
EasySep™ Mouse B Cell Isolation Kit
Catalog #
19854
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ Mouse B Cell Isolation Kit
Catalog #
19854RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ Mouse B Cell Isolation Kit
Catalog #
19854RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
RoboSep™ Mouse B Cell Isolation Kit
Catalog #
19854RF
Lot #
All
Language
English

Educational Materials(15)

Brochure
EasySep™ Cell Separation Technology
Brochure
Mouse Cell Isolation
Brochure
Tools For Your Immunology Research
Brochure
Cell Enrichment Prior to Cell Sorting
Wallchart
Antigen Processing and Presentation
Wallchart
Frequencies and Percentages of Mouse Immune Cell Types
Wallchart
Human Immune Cytokines
Video
0:51
EasySep™ Mouse Cell Isolation Kits: How Much Time Will You Save? (Part 2)
Video
3:58
Comparison of Mouse Immune Cell Isolation Protocols: EasySep™ vs. Column-Based Magnetic Separation
Video
0:52
EasySep™ Mouse Cell Isolation Kits: How Much Time Will You Save? (Part 3)
Video
0:01
EasySep™ Mouse Cell Isolation Kits: How Much Time Will You Save?
Video
1:13
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology
Video
1:57
How EasySep™ Magnetic Cell Separation Technology Works: Fast and Easy Cell Isolation
Video
0:49
EasySep™ Mouse Cell Isolation Kits: How Much Time Will You Save? (Part 1)
Video
0:57
Simultaneous Cell Isolation from Multiple Samples Using the EasyEights™ EasySep™ Magnet
Load More Educational Materials

Frequently Asked Question

Can EasySep™ Streptavidin RapidSpheres™ be used for either positive or negative selection?

Currently, EasySep™ Streptavidin RapidSphere™ kits are only available for negative selection and work by targeting and removing unwanted cells.

How does the separation work?

Streptavidin RapidSphere™ magnetic particles are crosslinked to unwanted cells using biotinylated antibodies. When placed in the EasySep™ Magnet, labeled cells migrate to the wall of the tube. The unlabeled cells are then poured off into a new tube.

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™ Streptavidin RapidSphere™ separations be automated?

Yes. RoboSep™, the fully automated cell separator, automates all EasySep™ labeling and cell separation steps.

Are cells isolated using EasySep™ RapidSphere™ products FACS-compatible?

Yes. Desired cells are unlabeled and ready to use in downstream applications, such as FACS analysis.

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.
Read More

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.

Data and Publications

Data

Typical EasySep™ Mouse B Cell Isolation Profile

Figure 1. Typical EasySep™ Mouse B Cell Isolation Profile

Starting with mouse splenocytes, the B cell content of the isolated fraction typically ranges from 94 - 98%.

Publications (30)

Nature biomedical engineering 2020 jun Pharmacokinetic tuning of protein-antigen fusions enhances the immunogenicity of T-cell vaccines. N. K. Mehta et al.

Abstract

The formulations of peptide-based antitumour vaccines being tested in clinical studies are generally associated with weak potency. Here, we show that pharmacokinetically tuning the responses of peptide vaccines by fusing the peptide epitopes to carrier proteins optimizes vaccine immunogenicity in mice. In particular, we show in immunized mice that the carrier protein transthyretin simultaneously optimizes three factors: efficient antigen uptake in draining lymphatics from the site of injection, protection of antigen payloads from proteolytic degradation and reduction of antigen presentation in uninflamed distal lymphoid organs. Optimizing these factors increases vaccine immunogenicity by up to 90-fold and maximizes the responses to viral antigens, tumour-associated antigens, oncofetal antigens and shared neoantigens. Protein-peptide epitope fusions represent a facile and generalizable strategy for enhancing the T-cell responses elicited by subunit vaccines.
Nature nanotechnology 2020 jun Role of nanoscale antigen organization on B-cell activation probed using DNA origami. R. Veneziano et al.

Abstract

Vaccine efficacy can be increased by arraying immunogens in multivalent form on virus-like nanoparticles to enhance B-cell activation. However, the effects of antigen copy number, spacing and affinity, as well as the dimensionality and rigidity of scaffold presentation on B-cell activation remain poorly understood. Here, we display the clinical vaccine immunogen eOD-GT8, an engineered outer domain of the HIV-1 glycoprotein-120, on DNA origami nanoparticles to systematically interrogate the impact of these nanoscale parameters on B-cell activation in vitro. We find that B-cell signalling is maximized by as few as five antigens maximally spaced on the surface of a 40-nm viral-like nanoparticle. Increasing antigen spacing up to {\~{}}25-30 nm monotonically increases B-cell receptor activation. Moreover, scaffold rigidity is essential for robust B-cell triggering. These results reveal molecular vaccine design principles that may be used to drive functional B-cell responses.
Scientific reports 2020 jul Bioluminescence for in vivo detection of cell-type-specific inflammation in a mouse model of uveitis. S. John et al.

Abstract

This study reports the use of cell-type-specific in vivo bioluminescence to measure intraocular immune cell population dynamics during the course of inflammation in a mouse model of uveitis. Transgenic lines expressing luciferase in inflammatory cell subsets (myeloid cells, T cells, and B cells) were generated and ocular bioluminescence was measured serially for 35 days following uveitis induction. Ocular leukocyte populations were identified using flow cytometry and compared to the ocular bioluminescence profile. Acute inflammation is neutrophilic (75{\%} of ocular CD45 + cells) which is reflected by a significant increase in ocular bioluminescence in one myeloid reporter line on day 2. By day 7, the ocular T cell population increases to 50{\%} of CD45 + cells, leading to a significant increase in ocular bioluminescence in the T cell reporter line. While initially negligible ({\textless} 1{\%} of CD45 + cells), the ocular B cell population increases to {\textgreater} 4{\%} by day 35. This change is reflected by a significant increase in the ocular bioluminescence of the B cell reporter line starting on day 28. Our data demonstrates that cell-type-specific in vivo bioluminescence accurately detects changes in multiple intraocular immune cell populations over time in experimental uveitis. This assay could also be useful in other inflammatory disease models.
Science advances 2020 B cell Sirt1 deacetylates histone and non-histone proteins for epigenetic modulation of AID expression and the antibody response. H. Gan et al.

Abstract

Activation-induced cytidine deaminase (AID) mediates immunoglobulin class switch DNA recombination (CSR) and somatic hypermutation (SHM), critical processes for maturation of the antibody response. Epigenetic factors, such as histone deacetylases (HDACs), would underpin B cell differentiation stage-specific AID expression. Here, we showed that NAD+-dependent class III HDAC sirtuin 1 (Sirt1) is highly expressed in resting B cells and down-regulated by stimuli inducing AID. B cell Sirt1 down-regulation, deprivation of NAD+ cofactor, or genetic Sirt1 deletion reduced deacetylation of Aicda promoter histones, Dnmt1, and nuclear factor-$\kappa$B (NF-$\kappa$B) p65 and increased AID expression. This promoted class-switched and hypermutated T-dependent and T-independent antibody responses or led to generation of autoantibodies. Genetic Sirt1 overexpression, Sirt1 boost by NAD+, or allosteric Sirt1 enhancement by SRT1720 repressed AID expression and CSR/SHM. By deacetylating histone and nonhistone proteins (Dnmt1 and NF-$\kappa$B p65), Sirt1 transduces metabolic cues into epigenetic changes to play an important B cell-intrinsic role in modulating antibody and autoantibody responses.
Nature communications 2020 B cell-intrinsic epigenetic modulation of antibody responses by dietary fiber-derived short-chain fatty acids. H. N. Sanchez et al.

Abstract

Short-chain fatty acids (SCFAs) butyrate and propionate are metabolites from dietary fiber's fermentation by gut microbiota that can affect differentiation or functions of T cells, macrophages and dendritic cells. We show here that at low doses these SCFAs directly impact B cell intrinsic functions to moderately enhance class-switch DNA recombination (CSR), while decreasing at higher doses over a broad physiological range, AID and Blimp1 expression, CSR, somatic hypermutation and plasma cell differentiation. In human and mouse B cells, butyrate and propionate decrease B cell Aicda and Prdm1 by upregulating select miRNAs that target Aicda and Prdm1 mRNA-3'UTRs through inhibition of histone deacetylation (HDAC) of those miRNA host genes. By acting as HDAC inhibitors, not as energy substrates or through GPR-engagement signaling in these B cell-intrinsic processes, these SCFAs impair intestinal and systemic T-dependent and T-independent antibody responses. Their epigenetic impact on B cells extends to inhibition of autoantibody production and autoimmunity in mouse lupus models.
Frontiers in immunology 2020 Recombinant Factor VIII Fc Inhibits B Cell Activation via Engagement of the Fc$\gamma$RIIB Receptor. M. T. Georgescu et al.

Abstract

The development of neutralizing antibodies (inhibitors) against factor VIII (FVIII) is a major complication of hemophilia A treatment. The sole clinical therapy to restore FVIII tolerance in patients with inhibitors remains immune tolerance induction (ITI) which is expensive, difficult to administer and not always successful. Although not fully understood, the mechanism of ITI is thought to rely on inhibition of FVIII-specific B cells (1). Its efficacy might therefore be improved through more aggressive B cell suppression. Fc$\gamma$RIIB is an inhibitory Fc receptor that down-regulates B cell signaling when cross-linked with the B cell receptor (BCR). We sought to investigate if recombinant FVIII Fc (rFVIIIFc), an Fc fusion molecule composed of FVIII and the Fc region of immunoglobulin G1 (IgG1) (2), is able to inhibit B cell activation more readily than FVIII. rFVIIIFc was able to bind FVIII-exposed and na{\{i}}ve B cells from hemophilia A mice as well as a FVIII-specific murine B cell hybridoma line (413 cells). An anti-Fc$\gamma$RIIB antibody and FVIII inhibited binding suggesting that rFVIIIFc is able to interact with both Fc$\gamma$RIIB and the BCR. Furthermore incubation of B cells from FVIII-exposed mice and 413 cells with rFVIIIFc resulted in increased phosphorylation of SH-2 containing inositol 5-phosphatase (SHIP) when compared to FVIII. B cells from FVIII-exposed hemophilia A mice also exhibited decreased extracellular signal-regulated kinase (ERK) phosphorylation when exposed to rFVIIIFc. These differences were absent in B cells from na{\"{i}}ve non-FVIII exposed hemophilic mice suggesting an antigen-dependent effect. Finally rFVIIIFc was able to inhibit B cell calcium flux induced by anti-Ig F(ab)2. Our results therefore indicate that rFVIIIFc is able to crosslink Fc$\gamma$RIIB and the BCR of FVIII-specific B cells causing inhibitory signaling in these cells."""
View All Publications

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