In a recent publication, researchers from the University of Miami Miller School of Medicine (USA) describe that Serotonin released by human beta cell inhibits glucagon secretion by alpha cells. They demonstrated that this paracrine loop was mediated via the cAMP pathway. To do so, they captured in live human pancreatic islet cells cAMP signals using a specific fluorescent biosensor.
Understanding how mammalian cells function requires a dynamic perspective. Recent improvements in our abilities to perform fluorescence microscopy on primary cells, coupled with advances in pipelines for quantifying and extracting data, have made possible a better understanding of the temporal complexity of cell signalling pathway. Due to the heterogeneity seen in both eukaryotic and prokaryotic cell populations, study at the single cell resolution with living cells is currently the best solution to understand the dynamics between environmental conditions and cellular behaviour.
However as living cells don’t stay still, classical imaging and studying method present some drawbacks for single cell analysis and tracking such as: [Read more…]
Many ELISA kits – one analysis tool
Analyte quantification (in pg/ml or ng/ml) such as IL-6, TNFalpha and numerous other proteins is conveniently performed with immunoassays called ELISA kits. At tebu-bio, we not only provide more than 10 000 ELISA kits but we also support researchers through our techserv team who are on-hand to suggest the best kits and to answer any questions.
Being part of that team, I have noticed a number of often asked questions that can be resumed in one : What is the best method for data analysis? [Read more…]
The Ras-Raf-MEK-ERK Pathway is an important cell signaling route with many implications for cancer biology and therapeutic development. Dis-regulation of some of this pathway’s proteins expression and phosphorylation status are observed in about one-third of all human cancers. Access to specific tools to study this pathway is essential to a better understanding of its role in cancer for novel drug development. If you are working on this topic, you’ll be interested in taking a look at the range of reagents on offer for a wide variety of applications.
Sequence engineering and chemical modification of CAS9 mRNA
For transgenesis and therapeutic perspectives, vector-free delivery of the CAS9 for gene editing is highly preferable. It avoids the risk of hitting the genome due to the vector integration. Directly using CAS9 mRNA is a top option.
Furthermore, as a new promising class of therapeutic biologics, messenger RNA are the subject of numerous studies.
This year, Matthew Porteus’s and Trilink’s teams revealed key and practical results at the ASGC with a poster you can download below.
Notably, there is a large screen of the mRNA CAS9 modifications based on the indel formation. This interesting comparison includes also CAS9 RNP and reveals the high performance of the CleanCap U-depleted 5-moU CAS9 mRNA. We can also note that for the CleanCap CAS9 mRNA HPLC purification is not required.
“Maximizing Translation of Cas9 mRNA Therapeutics by Sequence Engineering and Chemical Modification”
Download your copy of the poster here.
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Shen et al (Nature Methods, 2017) explore and identify synthetic interactions among 73 cancer-associated genes. To perform their loss of function screen they combined CAS9-expressing cell lines with a sgRNA library of high titer lentiviral particles. Most of these gene interactions were subsequently validated by drug treatment. [Read more…]
If you are looking for enzymatic in vitro tools for drug metabolism, reaction phenotyping, metabolite generation, using pig as model, you may find these newly released Pig Bactosomes from Cypex of interest.
Bactosomes are recombinant enzymes expressed in E. coli with a patented expression system that enables expression of drug metabolising enzymes without the need for large modifications to be made to the proteins. They demonstrate excellent batch-to-batch consistency and robust activity levels. Each batch is characterized for linearity with time and CYP concentration, Km and Vmax. [Read more…]
One of our aims is to offer you, the researcher, a large and varied selection of biologically relevant, innovative Live Cell Imaging tools. In 2016, we started a collaboration with Goryo Chemical, when our specialists selected their unique fluorescent dyes for cellular analysis, assays and live cell imaging.
In one of my previous posts, I already invited you to take a (non-exhaustive) look at their diversified range of fluorescent dyes, which can be used in a variety of fields such as stem cell differentiation, cancer cell studies, Reactive Oxygen Species (ROS) analysis… Today, I’d like to take you through two new fluorescent probes we’re pleased to make available for you.
Metabolite characterization can be a lengthy process making your in vitro drug testing time consuming and expensive. Whether you are working on phase I or phase II metabolic enzymatic reactions, take a look at these BMO kits which can help you speed up this characterisation process.
- Perform the primary screen and select the desired metabolite wells (XTHCK1001-01)
- Perform the optimization and identify the best production conditions
- Scale-up and isolate the metabolite
A single-domain antibody (sdAb, also called Nanobody) is an antibody fragment consisting of a single monomeric variable antibody domain. Like a whole antibody, it is able to bind selectively to a specific antigen. With a molecular weight of only 12–15 kDA, single-domain antibodies are much smaller than common antibodies (150–160 kDa). They have been shown to be just as specific as a regular antibody and in some cases more robust. They are being researched for multiple pharmaceutical applications in in vivo imaging and targeted therapy. eg. the fusion of a fluorescent protein to a single-domain antibody can be used to trace targets in different compartments of living cells. They can therefore increase the possibilities of live cell microscopy and will enable novel functional studies. [Read more…]