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.
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.
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Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
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Resources and Publications
Educational Materials (7)
Frequently Asked Questions
Publications (4)
Microfluidic Squeezing Enables MHC Class I Antigen Presentation by Diverse Immune Cells to Elicit CD8+ T Cell Responses with Antitumor Activity. 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. 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. 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.
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