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.
The 4 newly released pig Bacterial membranes contain the pig cytochrome P450s CYP1A2 (CYP452), CYP2C49 (CYP451), CYP2E1 (CYP454); CYP3A39 (CYP453), each co-expressed with pig NADPH-cytochrome P450 reductase.
The data shown below are taken from the CYP2E1 characterization as an example:
The CYP2E1 is high reductase supplemented with purified human cytochrome b5 to boost its activity activity. b5 was added to the prep to allow reduction of the Km for substrates (while with some other CYPs, b5 increases the Vmax without affecting the Km). A high reductase CYP3A39 supplemented with cytochrome b5 is to be released soon.
Should you want to receive more information on these enzymes, feel free to leave me a comment and I’ll be happy to reply.
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…]
We previously addressed the PD-1:PD-L1/PD-L2 immune inhibition and the corresponding cell-based tools to screen immuno-oncology agents for restoring antitumor immune response. Unfortunately cancer cells have other ways to escape and survive. Let look together at another immune inhibition pathway, namely the TIGIT:CD155/CD112.
As messenger RNAs (mRNAs) are easier to deliver into cells then plasmids or viral vectors, they are useful for non-dividing cells. Inversely to the vectors, they ensure genome integrity that is recommended for cell therapeutics. mRNA are also well adapted to transient expression as required for cellular reprogramming, gene editing, and vaccines.
You might like to take a look at this poster, presented by Trilink (renowned modified nucleic acid experts) at the Keystone symposia last March (Pattern Recognition Signaling: From Innate Immunity to Inflammatory Disease). They share results on how to optimize messenger RNA for therapeutic activity.
“Innate Immune focused approaches to maximize messenger RNA therapeutic activity”
Download your copy of the poster here.
The mitogen-activated protein kinase (MAPK) signaling pathway is activated by a number of extra and intracellular stimuli including cytokines, growth factors, and hormones as well as stressors such as oxidative and ER stress. This pathways plays a key role in the regulation of many cellular processes including proliferation, differentiation, the stress response, motility, growth, differentiation, survival, and death. Abnormal MAPK signaling may contribute to increased or uncontrolled cell proliferation and/or resistance to apoptosis. To study this complex pathway, several tools are available, from the pathway specific arrays for an initial screen, to phospho-specific ELISAs for individual target validation.
This post aims at helping you to easily identify tools to explore this pathway in your samples (from arrays to phospho-ELISAs). However, I could not start without showing you once more one of these pretty illustrations of cell signalling pathways. I’ll let you explore it to dig out the MAPK protein cascade among all of them (a kind of Where’s Wally for the researcher !).
Since the discovery of reprogramming factors in 2006 and the boom of CRISPR engineering strategies, iPSC cell lines have emerged as new cellular models. The development of 3D culture technologies has also contributed to the generation of iPSC derived cells, with unique applications from patient-specific drug responses testing, to regenerative medicine.
I would like to introduce in this post a selection of reagents in this domain, a combination of both routine and innovative quality reagents, that I consider as bringing something extra to your stem cell research project.
In a previous post dedicated to Quantitative arrays (Quantibody), I introduced our L-Series aimed at a broad one shot profiling of up to 1000 markers at once. This relative quantitation technology allows you to perform a first screen of your samples of interest versus a control, before you go on to targeted profiling using either pathway specific arrays, or a custom array including the targets of interest identified with the initial L-series screen. [Read more…]