Single-cell isolation allows genomic and transcriptomic analysis of one individual cell. It is also required to build a monoclonal cell line from rare cell isolation, that could be for example CRISPR-CAS9 gene edited cells. There are 3 popular methods: serial dilution, micro-manipulation and flow cytometry. None of these are easy or simple, and they often require expertise and experience. Fortunately, a new solution for everybody has come! It’s designed to isolate a single cell in a few seconds, and it’s (quite appropriately) called the Smart Aliquotor. [Read more…]
Cardiovascular disorders: Proteomics tool for disease signature identification
Cardiovascular disease (CVD) comprehensively describes a range of pathologies of the heart and/or blood vessels. These disorders differ markedly in time-scale of development, and relative contributory effects of genes and environment. Largely due to this breadth, and their causes and effects, the range of investigation, assessment, and treatment strategies available is large. The diversity of the diseases, disease processes, and genetic differences between individuals could obscure disease associations in terms of the proteomic signatures of each disorder. Thus, a panel of independent disease-related proteins considered together should be less prone to this noise and help provide much more robust analysis and indication of disease. The publication featured below and the reagents (multi-plex immunoassays or single target ELISA and antibodies) mentioned in this post may contribute in helping you to identify such diseases signature. [Read more…]
Tips and Tricks: how to increase unstable protein expression in E. Coli
Recombinant protein expression and purification for therapeutic development or for in vitro studies can sometimes be a real challenge. It requires huge investments in both time and cost in order to obtain high yields of pure and active recombinant proteins. My aim in writing this post, is to give you a simple guide to easily establish optimal conditions for recombinant protein expression and production in E. Coli, of problematic unstable proteins of interest.
Kupffer cells in co-culture models for liver inflammation studies
Kupffer primary cells are macrophages endogenous to the liver which have the ability to modulate hepatic inflammation and injury associated with various pathophysiologies and toxicities. Pro-inflammatory cytokines released by activated Kupffer cells, such as TNF-α and IL-6, are associated with up-regulation of acute-phase response proteins and suppression of CYP enzymes. For new biological entities, particularly immunomodulators, evaluating the potential for Kupffer cell activation is an emerging concept in preclinical development. Kupffer cells are estimated to comprise approximately 4-8% of total liver cell content and approximately 20% of non-parenchymal cells (Raccanelli and Rehermann, 2006). [Read more…]
Cell signalling – Discover G-LISA to study Small GTPases
As is already known, multiple families of small GTPases (Ras, Arf and Rho families) make up the Ras 
GTPase superfamily. Due to their role as molecular switches in cell signalling (active GTP-bound state vs inactive GDP-bound states) and in many other cellular responses (cytoskeletal reorganization, regulation of transcription, apoptosis…), these small GTPases are the subject of intense investigation to try to understand the mechanisms that regulate activation and inactivation of this proteins.
In this post, I invite you to discover a revolutionary way to study these cell signalling events by quantifying the level of active small GTPases in your experimental model.
Neurons, Astrocytes, Microglia: High Quality primary cells source
In the field of neurosciences, increasing research is being undertaken on neurodegenerative disorders like Parkinson’s and Alzheimer’s diseases, as well as on nerve injury and regeneration. One of the challenges researchers face is finding the appropriate cellular neuromodels, as close as possible to the in vivo environment. Primary cells are one of the answers, increasingly used in co-culture systems and 3D models. However, cells from the nervous system are not readily accessible, nor are they easy to culture.
Cell Applications Inc. provide solutions to face these challenges, ranging from highly pure, low passage ready-to-use primary cells, to stem cells and induced pluripotent stem cells (iPSC) derived from a variety of clinically relevant species. Let’s take a look at a selection of their neuron, microglia and astrocyte models in relation to their potential applications.
[Read more…]
Cancer immunotherapy: explore the cytokine “milieu” with Quantibody 200 Biomarker Service
Autologous tumour cell vaccines vs. dendritic cell vaccines: What’s best, and why?
Understanding the best approach for tumour vaccinations is of growing importance in the personalized cancer immunotherapy field. Current models use a variety of treatments, but two include the use of irradiated autologous tumour cells (TCV), or autologous dendritic cells loaded with tumour associated antigens (DCV). In a randomized trial of metastatic melanoma patients undergoing either TCV or DCV, DCV patient survival was far superior at 43.4 vs 20.5 months. Why was the response more effective when the same antigens were provided to the immune response? Dillman, Robert O., et al. brought some answers in a paper published in the Journal for immunotherapy of cancer in 2018). Looking at the cytokine “milieu” with the Quantibody 200 Biomarker Microarray Service helped the authors in understanding the differences in vaccine responses. [Read more…]
HDACs: Active drug targets for Immunotherapy treatment
For many pathologies such as cancer or neurological disorders, it has been found that Histone Deacetylase (HDAC) activity is disrupted, making them promising targets for drug development. Scientists need access to a broade range of products ( Active Enzyme, inhibitors/activators, Biochemical screening assays…), to simplify their research and identification of HDAC inhibitors.
Antibody reproducibility: what will make the difference…
Rockland Immunochemicals, Inc., (Rockland) provide the highest quality antibodies to the academic, biopharma, and diagnostic industries for use in basic research, assay development, preclinical and clinical studies, and bioprocessing. With facilities in Pennsylvania established since 55 years ago, Rockland manufactures products ideally suited for integration into critical assays such as western blotting, immunohistochemistry (IHC), immunofluorescence microscopy (IF), ELISA, flow cytometry, and 2D imaging. Their antibodies show remarkable quality and reproducibility from batch to batch. Their popular anti-RFP is one example out of many.
Demonstrating Rockland’s commitment in this matter (manufacturing antibodies of high quality and reproducibility), Rockland’s CSO, Dr. Carl Ascoli, testified at the US National Academies of Sciences, Engineering and Medicine’s third of six public sessions on Reproducibility and Replicability in Science on April 18th this year. [Read more…]
Maximize metabolic activities by limiting Hepatocyte cryoinjury
I am following up on my series of posts based on the work done by Chris Bohl and collaborators at Sekisui Xenotech. Here, he looks at the effect of cryopreservation on pooled hepatocytes metabolic activities, and how the patented Cryostax technology can address this issue.