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 include a Mouse FcR blocker instead of Normal Rat Serum. With this change, all components will now be shipped in a single package, while providing the same cell isolation performance as before.
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Easily and efficiently isolate highly purified mouse monocytes from mouse bone marrow, splenocytes, whole blood, or other single-cell suspension samples by immunomagnetic negative selection, with the EasySep™ Mouse Monocyte 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. The following unwanted cells are targeted for removal: granuloctyes, T cells, B cells, NK cells, hematopoietic progenitor cells, and erythroid cells. The magnetically labeled cells are then separated from the untouched desired monocytes 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 15 minutes, the desired monocytes are ready for downstream applications such as flow cytometry, culture, or cell-based assays.
Learn more about how immunomagnetic EasySep™ technology works or how to fully automate immunomagnetic cell isolation with Dzdz™. Explore additional products optimized for your workflow, including culture media, supplements, antibodies, and more.
Starting with mouse bone marrow cells, the monocyte content (Lineage- (CD3, CD45R, CD117, CD49b, Siglec F) CD11b+Ly6G- Ly6Chi/lo) of the isolated fraction is 89.5 ± 4.8% (mean ± SD), using the purple EasySep™ Magnet. In the above example, monocyte purities in the start and final isolated fractions are 7.1% and 92.3%, respectively.
Figure 2. Data for Anti-Mouse CD11b Antibody, Clone M1/70, Alexa Fluor® 488-Conjugated
(A) Flow cytometry analysis of C57BL/6 mouse splenocytes labeled with Anti-Mouse CD11b Antibody, Clone M1/70, Alexa Fluor® 488 (Catalog #60001AD) and anti-mouse CD45 APC. (B) Flow cytometry analysis of C57BL/6 mouse splenocytes labeled with a rat IgG2b, kappa Alexa Fluor® 488 isotype control antibody and anti-mouse CD45 APC. (C) Flow cytometry analysis of C57BL/6 mouse splenocytes processed with the EasySep™ Mouse Monocyte Enrichment Kit (Catalog #19861) and labeled with Anti-Mouse CD11b Antibody, Clone M1/70, Alexa Fluor® 488 (Catalog #60001AD). Histograms show labeling of splenocytes (Start) and isolated cells (Isolated). Labeling of start cells with a rat IgG2b, kappa Alexa Fluor® 488 isotype control antibody is shown in the bottom panel (open histogram). (D) Flow cytometry analysis of C57BL/6 mouse bone marrow cells processed with the EasySep™ Mouse Monocyte Enrichment Kit (Catalog #19861) and labeled with Anti-Mouse CD11b Antibody, Clone M1/70, Alexa Fluor® 488 (Catalog #60001AD). Histograms show labeling of bone marrow cells (Start) and isolated cells (Isolated). Labeling of start cells with a rat IgG2b, kappa Alexa Fluor® 488 isotype control antibody is shown in the bottom panel (open histogram).
Figure 3. Cell Isolation Protocol Lengths
Typical time taken (in minutes) to isolate cells using select EasySep™ kits.
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.
Enhanced resistance to Listeria infection in mice surviving sepsis: the role of lipid metabolism and myeloid cell reprogramming
Frontiers in Pharmacology 2025 May
Abstract
IntroductionImmune resilience is the capacity of the immune system to recover from sepsis-induced organ injury and reestablish host defense. While sepsis survivors are often viewed as immunocompromised, recent studies suggest that some may acquire adaptive immune traits that enhance resistance to secondary infections.MethodsWe employed a murine cecal ligation and puncture (CLP) model to study polymicrobial sepsis and subsequent immune responses. Listeria monocytogenes was used as a model intracellular pathogen to assess immune protection. We analyzed myeloid cell phenotypes using single-cell RNA sequencing and evaluated lipid metabolic changes through quantitative RT-PCR, immunohistochemistry, serum cytokine assays, and plasma lipidomics.ResultsSepsis-surviving mice showed enhanced resistance to Listeria infection. Single-cell RNA sequencing revealed transcriptional reprogramming in splenic CD11b+Ly6Chigh myeloid cells, including downregulation of lipoprotein lipase and lipid efflux genes. CD11b+ myeloid cells exhibited increased lipid droplet accumulation, accompanied by elevated serum interferon-gamma (IFN-γ) levels. Plasma lipidomics identified systemic lipid remodeling, with increased phosphatidylserine and decreased phosphatidylinositol and phosphatidylglycerol levels.DiscussionOur findings suggest that sepsis survival induces lipid metabolic reprogramming in myeloid cells, contributing to enhanced immunity against intracellular pathogens. These insights reveal potential therapeutic targets within lipid metabolic pathways to improve host defense in sepsis survivors.
H3K27me3 modulates trained immunity of monocytes in HDM-allergic diseases
Frontiers in Immunology 2025 May
Abstract
BackgroundMonocytes have been confirmed to increase in persistently food-allergic children. A phenomenon of innate immune memory, called trained immunity, has also been observed in monocytes from allergic children. However, the underlying mechanism remains poorly understood.MethodsWe enrolled a cohort of HDM-allergic children alongside age-matched healthy controls and established an HDM-sensitized allergic mouse model. Flow cytometric analyses were conducted to quantify monocyte frequencies in clinical cohorts and experimental animals. We performed integrated transcriptomic profiling via RNA-seq combined with chromatin occupancy analysis using CUT&Tag technology in parallel human and murine samples to elucidate the molecular mechanisms.ResultsIn our study, we demonstrated a reduced H3K27me3 methylation level accompanied by an increased proportion and a proinflammatory transcriptional memory in monocytes from house dust mite (HDM)-allergic human subjects. The same transcriptional and epigenetic phenotype was also confirmed in HDM-sensitized mice. Finally, the administration of GSK-J4, which upregulates H3K27me3 level in murine monocytes, attenuated the inflammatory response in vitro and in vivo.ConclusionsOur study confirms that H3K27me3 methylation modulates the trained immunity in monocytes and regulates HDM-allergic diseases through an inflammatory-dependent mechanism.
KLF family members control expression of genes required for tissue macrophage identities
The Journal of Experimental Medicine 2025 Mar
Abstract
This work demonstrates that group 2 KLF family transcription factors are critical for specifying the identity of distinct tissue-resident macrophages. KLF2 directly controls expression of genes previously shown to be necessary in cavity macrophages, while KLF4 may play a similar role in alveolar macrophages. Tissue-resident macrophages adopt distinct gene expression profiles and exhibit functional specialization based on their tissue of residence. Recent studies have begun to define the signals and transcription factors that induce these identities. Here we describe an unexpected and specific role for the broadly expressed transcription factor Krüppel-like factor 2 (KLF2) in the development of embryonically derived large cavity macrophages (LCMs) in the serous cavities. KLF2 not only directly regulates the transcription of genes previously shown to specify LCM identity, such as retinoic acid receptors and GATA6, but also is required for induction of many other transcripts that define the identity of these cells. Our results suggest that KLF4 may similarly regulate the identity of alveolar macrophages in the lung. These data demonstrate that broadly expressed transcription factors, such as group 2 KLFs, can play important roles in the specification of distinct identities of tissue-resident macrophages.
(C3H x BALB/c) F1 hybrid monoclonal IgG2a antibody against mouse NK1.1 (CD161)
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EasySep™ Mouse Monocyte 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 include a Mouse FcR blocker instead of Normal Rat Serum. With this change, all components will now be shipped in a single package, while providing the same cell isolation performance as before.
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 ƽ, REFER TO WWW.ƽ.COM/COMPLIANCE.