EasySep™ Mouse Pan-B Cell Isolation Kit

Immunomagnetic negative isolation of untouched mouse pan-B (CD19+, CD19+CD138+, CD138+) cells

New format, same high quality! You may notice that your kit contents and packaging look slightly different from previous orders. We are currently updating the format of select EasySep™ Mouse kits to remove Normal Rat Serum, as this has been found to improve cell isolation performance. With this change, all components will now be shipped in a single package.

EasySep™ Mouse Pan-B Cell Isolation Kit

Immunomagnetic negative isolation of untouched mouse pan-B (CD19+, CD19+CD138+, CD138+) cells

From: 839 USD
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Immunomagnetic negative isolation of untouched mouse pan-B (CD19+, CD19+CD138+, CD138+) cells
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Product Advantages


  • Fast and easy-to-use

  • Up to 98% purity

  • No columns required

  • Untouched, viable cells

What's Included

  • EasySep™ Mouse Pan-B Cell Isolation Kit (Catalog #19844)
    • EasySep™ Mouse Pan-B Cell Isolation Cocktail, 0.5 mL
    • EasySep™ Streptavidin RapidSpheres™ 50001, 1 mL
  • RoboSep™ Mouse Pan-B Cell Isolation Kit (Catalog #19844RF)
    • EasySep™ Mouse Pan-B Cell Isolation Cocktail, 0.5 mL
    • EasySep™ Streptavidin RapidSpheres™ 50001, 1 mL
    • RoboSep™ Buffer (Catalog #20104)
    • RoboSep™ Filter Tips (Catalog #20125)

Overview

Easily and efficiently isolate highly purified mouse pan-B cells (CD19+, CD19+CD138+, and CD138+), including conventional B-2 B cells, B-1 B cells and plasma cells from single-cell suspensions of splenocytes or other tissue samples by immunomagnetic negative selection, with the EasySep™ Mouse Pan-B Cell Isolation Kit. 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: CD4, CD8, CD11c, CD49b, CD90.2, Ly-6C/G (Gr-1) and TER119. The magnetically labeled cells are then separated from the untouched desired pan-B cells by using an EasySep™ magnet and simply pouring or pipetting the desired cells into a new tube. Following magnetic cell isolation in as little as 10 minutes, the desired pan-B cells are ready for downstream applications such as flow cytometry, culture, or cell-based assays.

This kit is compatible for use with cells from disease models where the malignant cells (B-CLL) express CD43 or CD11b. For isolation of conventional B cells only, we recommend using the EasySep™ Mouse B Cell Isolation Kit (Catalog #19854).

Learn more about how immunomagnetic EasySep™ technology works or how to fully automate immunomagnetic cell isolation with RoboSep™. Explore additional products optimized for your workflow, including culture media, supplements, antibodies, and more.
Magnet Compatibility
• EasySep™ Magnet (Catalog #18000)
• “The Big Easy” EasySep™ Magnet (Catalog #18001)
• 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

Data Figures

Typical EasySep™ Mouse Pan-B Cell Isolation Profile of a Non-Immunized C57BL/6 Mouse

Figure 1. Typical EasySep™ Mouse Pan-B Cell Isolation Profile of a Non-Immunized C57BL/6 Mouse

Starting with mouse splenocytes, the pan-B cell content (CD19+, CD19+CD138+, and CD138+) of the isolated fraction typically ranges from 91 - 98%.

Typical EasySep™ Mouse Pan-B Cell Isolation Profile of an Immunized C57BL/6 Mouse

Figure 2. Typical EasySep™ Mouse Pan-B Cell Isolation Profile of an Immunized C57BL/6 Mouse

Starting with mouse splenocytes, the pan-B cell content (CD19+, CD19+CD138+ and CD138+) of the isolated fraction typically ranges from 91 - 98%.

Protocol Diagram for Culturing Mouse B Cells with ImmunoCult™ Mouse B Cell Expansion Kit

Figure 3. Protocol Diagram for Culturing Mouse B Cells with ImmunoCult™ Mouse B Cell Expansion Kit

B cells isolated from mouse spleen using EasySep™ Mouse Pan-B Cell Isolation Kit (Catalog #19844) were cultured in complete Mouse B Cell Expansion Medium (Catalog #100-1004) as described in the Directions for Use (steps 1 - 7) of the PIS for ImmunoCult™ Mouse B Cell Expansion Kit (Catalog # 100-1003). B Cells were harvested on Day 9 for analysis. Cells can also be harvested at earlier time points depending on different applications.

Expansion of Mouse B Cells with ImmunoCult™ Mouse B Cell Expansion Kit

Figure 4. Expansion of Mouse B Cells with ImmunoCult™ Mouse B Cell Expansion Kit

B cells isolated from mouse spleen using EasySep™ Mouse Pan-B Cell Isolation Kit (Catalog #19844) were cultured in complete Mouse B Cell Expansion Medium (Catalog #100-1004) as described in PIS for ImmunoCult™ Mouse B Cell Expansion Kit (Catalog # 100-1003). Fold expansion of viable cells is shown with bar graphs representing the mean ± SEM (n = 8). B cells expanded 176.9 ± 29.8-fold after 9 days of culture.

Maturation of Mouse B Cells with ImmunoCult™ Mouse B Cell Expansion Kit

Figure 5. Maturation of Mouse B Cells with ImmunoCult™ Mouse B Cell Expansion Kit

B cells isolated from mouse spleen using EasySep™ Mouse Pan-B Cell Isolation Kit (Catalog #19844) were cultured in complete Mouse B Cell Expansion Medium (Catalog #100-1004) as described in the PIS for ImmunoCult™ Mouse B Cell Expansion Kit (Catalog # 100-1003). Following staining using the protocol by Pracht et al. (Eur J Immunol, 2017), the expression of A) B220 and CD138 and B) TACI (CD267) and CD86 were analyzed by flow cytometry at several time points (data represents mean ± SEM, n = 8). An increase in CD86 cell surface expression indicates B cell activation; a decrease in B220 and an increase in CD138 and TACI cell surface expression indicate maturation of B cells to plasmablasts or plasma cells.

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 #
19844
Lot #
1000147362 or lower
Language
English
Catalog #
19844
Lot #
1000147363 or higher
Language
English
Catalog #
19844RF
Lot #
1000147362 or lower
Language
English
Catalog #
19844RF
Lot #
1000147363 or higher
Language
English
Document Type
Safety Data Sheet 1
Catalog #
19844
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
19844
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Catalog #
19844
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
19844RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
19844RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Catalog #
19844RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Catalog #
19844RF
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™ 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.

Publications (4)

Microfluidic Squeezing Enables MHC Class I Antigen Presentation by Diverse Immune Cells to Elicit CD8+ T Cell Responses with Antitumor Activity. M. G. Booty et al. Journal of immunology (Baltimore, Md. : 1950) 2022 feb

Abstract

CD8+ T cell responses are the foundation of the recent clinical success of immunotherapy in oncologic indications. Although checkpoint inhibitors have enhanced the activity of existing CD8+ T cell responses, therapeutic approaches to generate Ag-specific CD8+ T cell responses have had limited success. Here, we demonstrate that cytosolic delivery of Ag through microfluidic squeezing enables MHC class I presentation to CD8+ T cells by diverse cell types. In murine dendritic cells (DCs), squeezed DCs were ˆ¼1000-fold more potent at eliciting CD8+ T cell responses than DCs cross-presenting the same amount of protein Ag. The approach also enabled engineering of less conventional APCs, such as T cells, for effective priming of CD8+ T cells in vitro and in vivo. Mixtures of immune cells, such as murine splenocytes, also elicited CD8+ T cell responses in vivo when squeezed with Ag. We demonstrate that squeezing enables effective MHC class I presentation by human DCs, T cells, B cells, and PBMCs and that, in clinical scale formats, the system can squeeze up to 2 billion cells per minute. Using the human papillomavirus 16 (HPV16) murine model, TC-1, we demonstrate that squeezed B cells, T cells, and unfractionated splenocytes elicit antitumor immunity and correlate with an influx of HPV-specific CD8+ T cells such that >80% of CD8s in the tumor were HPV specific. Together, these findings demonstrate the potential of cytosolic Ag delivery to drive robust CD8+ T cell responses and illustrate the potential for an autologous cell-based vaccine with minimal turnaround time for patients.
Quantitative and Qualitative Analysis of Humoral Immunity Reveals Continued and Personalized Evolution in Chronic Viral Infection. N. J. Kr\autler et al." Cell reports 2020 jan

Abstract

Control of established chronic lymphocytic choriomeningitis virus (LCMV) infection requires the production of neutralizing antibodies, but it remains unknown how the ensemble of antibodies evolves during ongoing infection. Here, we analyze the evolution of antibody responses during acute or chronic LCMV infection, combining quantitative functional assays and time-resolved antibody repertoire sequencing. We establish that antibody responses initially converge in both infection types on a functional and repertoire level, but diverge later during chronic infection, showing increased clonal diversity, the appearance of mouse-specific persistent clones, and distinct phylogenetic signatures. Chronic infection is characterized by a longer-lasting germinal center reaction and a continuous differentiation of plasma cells, resulting in the emergence of higher-affinity plasma cells exhibiting increased antibody secretion rates. Taken together, our findings reveal the emergence of a personalized antibody response in chronic infection and support the concept that maintaining B cell diversity throughout chronic LCMV infection correlates with the development of infection-resolving antibodies.
Genetically Engineered Cell-Derived Nanoparticles for Targeted Breast Cancer Immunotherapy. X. Shi et al. Molecular therapy : the journal of the American Society of Gene Therapy 2019 nov

Abstract

Exosomes are nanosized membranous vesicles secreted by a variety of cells. Due to their unique and pharmacologically important properties, cell-derived exosome nanoparticles have drawn significant interest for drug development. By genetically modifying exosomes with two distinct types of surface-displayed monoclonal antibodies, we have developed an exosome platform termed synthetic multivalent antibodies retargeted exosome (SMART-Exo) for controlling cellular immunity. Here, we apply this approach to human epidermal growth factor receptor 2 (HER2)-expressing breast cancer by engineering exosomes through genetic display of both anti-human CD3 and anti-human HER2 antibodies, resulting in SMART-Exos dually targeting T cell CD3 and breast cancer-associated HER2 receptors. By redirecting and activating cytotoxic T cells toward attacking HER2-expressing breast cancer cells, the designed SMART-Exos exhibited highly potent and specific anti-tumor activity both in vitro and in vivo. This work demonstrates preclinical feasibility of utilizing endogenous exosomes for targeted breast cancer immunotherapy and the SMART-Exos as a broadly applicable platform technology for the development of next-generation immuno-nanomedicines.
New format, same high quality! You may notice that your kit contents and packaging look slightly different from previous orders. We are currently updating the format of select EasySep™ Mouse kits to remove Normal Rat Serum, as this has been found to improve cell isolation performance. With this change, all components will now be shipped in a single package.