EasySep™ Human CD14 Positive Selection Kit II
Immunomagnetic positive selection of human CD14+ cells
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Protocols and Documentation.
- EasySep™ Magnet
Magnet for column-free immunomagnetic separation
- EasySep™ Buffer
Cell separation buffer for use with EasySep™ cell separation kits
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Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
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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.
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Educational Materials (9)
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Publications (43)
A semi‐automated ASC speck assay to evaluate pyrin inflammasome activation
Clinical & Translational Immunology 2025 Oct
Abstract
Objective: To develop a rapid functional assay to validate variants of uncertain significance (VUS) in the MEFV gene. Methods: Overactivity of the pyrin inflammasome pathway and ASC speck oligomerisation in response to stimulation with low concentrations of Clostridium difficile toxin A was directly visualised by immunofluorescence microscopy. A semi‐automated algorithm was developed to count cells and ASC specks. Results: The semi‐automated ASC speck assay is able to discriminate between healthy controls and patients with familial Mediterranean fever (FMF) and pyrin inflammasome overactivity with high sensitivity. It is also able to discriminate pyrin inflammasome overactivity from other autoinflammatory disease controls with high specificity. Conclusion: The semi‐automated ASC speck assay may be a useful test to functionally validate VUS in the MEFV gene and screen for pyrin inflammasome overactivity. A semi‐automated ASC speck assay using machine learning is able to discriminate between healthy controls and patients with familial Mediterranean fever (FMF) with high sensitivity. It is also able to discriminate FMF from other autoinflammatory diseases with high specificity.
Identification and characterization of a ubiquitin E3 RING ligase of the Chlamydia-like bacterium Simkania negevensis
PLOS Pathogens 2025 Nov
Abstract
In the arms race between a pathogen and the host, the defense mechanisms of the host cell, including the ubiquitin system, are often counteracted by bacteria. Simkania negevensis (Sne), an obligate intracellular Chlamydia-like bacterium connected with respiratory diseases, possesses numerous deubiquitinases, but not much is known about its other ubiquitin-modifying enzymes. Sne infects a wide range of hosts, developing inside a tubular vacuole in close contact with the host endoplasmic reticulum (ER) and mitochondria. Our study describes an uncharacterized Sne ubiquitin E3 RING-ligase (SNE_A12920 or SneRING), which primarily generates K63- and K11-linked ubiquitin chains and preferentially interacts with UbcH5b and UBE2T E2 enzymes. SneRING is expressed upon infection of various human cell lines, as well as amoebae. We show that a portion of the expressed SneRING co-localizes with mitochondria and ER and that the SneRING interactome includes mitochondrial and ER proteins involved in organelle morphology and stress response. Our work offers an initial characterization of a bacterial RING ligase potentially involved in the host cell remodeling to accommodate the unique intracellular lifestyle of Sne. Author summaryUbiquitination is a protein modification system that regulates protein degradation, localization, or interactions. As such, ubiquitination has many important functions in cell signalling, and its dysregulation can lead to cancer and neurodegenerative diseases. Bacteria that live and develop inside human or other eukaryotic cells, such as Chlamydia, often modulate the ubiquitination system to ensure their own survival. Simkania negevensis is a Chlamydia-like bacterium connected to respiratory diseases in humans. We have discovered a novel enzyme expressed by these bacteria that can ubiquitinate other proteins and thus potentially modify host cell processes that would otherwise hinder infection. In this work, we explore the function of this enzyme and determine its possible cellular localization, as well as some of the proteins it interacts with. Our study provides new insights into how bacterial pathogens adapt to and manipulate host cells using one of the major cell function regulatory systems.
Selection and Characterisation of Minor Histocompatibility Antigen‐Specific Regulatory T Cells in Fully HLA‐Matched Setting for GVHD Therapy
European Journal of Immunology 2025 Dec
Abstract
Graft‐versus‐host disease is mediated by donor‐derived T cells reactive against the recipient's broadly expressed minor histocompatibility antigens (mHA). Regulatory T cells (Treg) have been explored as a therapeutic approach for chronic GVHD (cGVHD). The promising results from polyclonal Treg trials in this setting have led us to develop a Treg product specific for mismatched minor antigens between patient and donor (mTreg), circumventing broad immune suppression risks. HLA‐matched siblings of opposite sexes were used to obtain the sister's CD4+CD25hiCD127low Treg for co‐culture with the respective brother's dendritic cells as a source of mismatched mHA. We have established the optimal culture conditions resulting in the highest mTreg proliferation and viability. Comprehensive phenotyping during the ex vivo selection shows PD‐1, CTLA‐4, CD39 and HLA‐DR expression. Transcriptomic analysis revealed a switch in metabolic process, and up‐regulation of functional Treg genes. Furthermore, mTreg possess specific and potent suppressive activity, in which there is a dependency on cell‐to‐cell contact and a role for HLA class II expression on mTreg. This protocol would allow the generation of Treg specific to an array of mHA from the recipient's healthy tissues, likely providing a directed and strong suppression of cGVHD. We optimised a protocol for mHA‐specific Treg (mTreg) selection in an HLA‐matched context while defining its phenotype, transcriptional state and function. mTreg were highly activated and exerted specific, HLA class II‐, contact‐dependent suppression. This protocol can be explored as a highly personalised antigen‐specific Treg‐based therapy in future clinical trials for cGVHD.
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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.
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.
