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.
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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.
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%.
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%.
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.
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.
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.
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.
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.
Differential metabolic pathways underlie THC- and CBD-mediated inhibition of B-cell activation in both young and aged mice
Frontiers in Immunology 2025 Jun
Abstract
ObjectiveB lymphocytes play a crucial role in immunity but also contribute to the pathogenesis of various diseases. Cannabis plants produce numerous biologically active compounds, including cannabinoids. The two most studied phytocannabinoids are Δ9-tetrahydrocannabinol (THC) and cannabidiol (CBD). These cannabinoids exert diverse and potent biological effects primarily through the endocannabinoid system (ECS), which also plays a key role in mature B-cell function. Both the immune system and the ECS undergo age-related changes that lead to a clinically significant decline in function.MethodsThis study compares the effects of THC and CBD on B-cell activity in young and aged mice. Murine B lymphocytes were activated using lipopolysaccharide (LPS) and interleukin-4 (IL-4), and the impact of cannabinoid treatments was assessed in terms of cell phenotype, proliferation, antibody secretion, tumor necrosis factor-alpha (TNFα) secretion, extracellular signal-regulated kinase (ERK) phosphorylation, and the cellular metabolome.ResultsBoth THC and CBD exhibited dose-dependent inhibitory effects on B-cell activation in young and aged mice. However, we show here, for the first time, that the treatments induce distinct metabolic profiles. Although some metabolites, such as glucose-6-phosphate, pentose phosphate pathway (PPP) and nucleotide metabolites, were reduced by both cannabinoids, THC selectively reduced the levels of a distinct set of amino acids, while only CBD increased the levels of Citrulline and Allantoin. Additionally, the effects of THC and CBD differed between young and aged B cells, suggesting that age-related changes in the ECS may influence cannabinoid sensitivity.ConclusionsThese findings provide insights into the distinct mechanisms by which THC and CBD regulate immune activation and may open the door for investigating the mechanisms behind cannabinoids effects on the immune system. They also highlight the need for further research into phytocannabinoid-based therapies, particularly in age-specific contexts. Given the immunoregulatory properties of cannabinoids, especially CBD, tailored therapeutic strategies may enhance their clinical applications
Longitudinal omics data and preclinical treatment suggest the proteasome inhibitor carfilzomib as therapy for ibrutinib-resistant CLL
Nature Communications 2025 Jan
Abstract
Chronic lymphocytic leukemia is a malignant lymphoproliferative disorder for which primary or acquired drug resistance represents a major challenge. To investigate the underlying molecular mechanisms, we generate a mouse model of ibrutinib resistance, in which, after initial treatment response, relapse under therapy occurrs with an aggressive outgrowth of malignant cells, resembling observations in patients. A comparative analysis of exome, transcriptome and proteome of sorted leukemic murine cells during treatment and after relapse suggests alterations in the proteasome activity as a driver of ibrutinib resistance. Preclinical treatment with the irreversible proteasome inhibitor carfilzomib administered upon ibrutinib resistance prolongs survival of mice. Longitudinal proteomic analysis of ibrutinib-resistant patients identifies deregulation in protein post-translational modifications. Additionally, cells from ibrutinib-resistant patients effectively respond to several proteasome inhibitors in co-culture assays. Altogether, our results from orthogonal omics approaches identify proteasome inhibition as potentially attractive treatment for chronic lymphocytic leukemia patients resistant or refractory to ibrutinib. The molecular mechanisms underlying resistance to therapy in Chronic lymphocytic leukemia (CLL) remain to be explored. Here, the authors perform multi-omics analysis in a mouse model of ibrutinib resistance and suggest proteasome inhibition for overcoming it.
Integrative multi-omics reveals a regulatory and exhausted T-cell landscape in CLL and identifies galectin-9 as an immunotherapy target
Nature Communications 2025 Aug
Abstract
T-cell exhaustion contributes to immunotherapy failure in chronic lymphocytic leukemia (CLL). Here, we analyze T cells from CLL patients’ blood, bone marrow, and lymph nodes, as well as from a CLL mouse model, using single-cell RNA sequencing, mass cytometry, and tissue imaging. T cells in CLL lymph nodes show the most distinct profiles, with accumulation of regulatory T cells and CD8+ T cells in various exhaustion states, including precursor (TPEX) and terminally exhausted (TEX) cells. Integration of T-cell receptor sequencing data and use of the predicTCR classifier suggest an enrichment of CLL-reactive T cells in lymph nodes. Interactome studies reveal potential immunotherapy targets, notably galectin-9, a TIM3 ligand. Inhibiting galectin-9 in mice reduces disease progression and TIM3+ T cells. Galectin-9 expression also correlates with worse survival in CLL and other cancers, suggesting its role in immune evasion and potential as a therapeutic target. Multi-omics can be used to characterise tumour and immune cell populations. Here the authors use multi-omics to characterise CLL blood and tissue samples and use prediction models for CLL TCR specificity and implicate interactions between galectin-9 and TIM3 as involved in CLL immune escape and propose galectin-9 as a possible immunotherapy target.
Rat monoclonal IgG2a antibody against mouse CD138 (syndecan-1)
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EasySep? Mouse Pan-B Cell Isolation Kit
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.
Quality Statement:
PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT 海角破解版, REFER TO WWW.海角破解版.COM/COMPLIANCE.