Nowadays, especially through the advent of systems biology, and the ability to generate predictive models, it is even more important to transform biology from a qualitative to a quantitative science. Interactions between proteins not only need to be identified, but their equilibrium rate constants need to be determined as well. Here we describe a very elegant and simple system to obtain quantitative data for protein-protein interactions using cell-free protein biosynthesis in combination with Microscale Thermophoresis. We have been able to characterize the interaction of Calmodulin with Ca2+ as well as its ligand M13 straight in the in vitro synthesis reaction. Furthermore, we are demonstrating the generic applicability of this approach with another set of proteins, an Antibody fragment and Cyclophilin A. All this work has been done without tedious expression and prior purification of the interacting proteins.
Have a question about Monolith?
Contact SpecialistSee more related content
43:09Learn about technologies and instruments that maintain membrane protein stability and allow structural abd affinity screening studies in their natural lipid environment.
Achieve Highly Sensitive and Precise Binding Affinity Measurements Without Disrupting Protein Conformation
15:19Virtual Monolith X Demo: Free-in-solution affinity measurements with minimal sample consumption
The tool you choose to measure affinities should tackle a diverse range of targets and ligands that come your way — such as GPCRs, IDPs, or cancer-related proteins. And, you should get data that help
20:31The power of Monolith X for Characterizing GPCRs: Accelerating Binding Affinity Measurements
To understand signaling pathways, Spectral Shift technology allows for accurate bindings affinity measurements of membrane proteins like GPCRs in close to native conditions.
Three things we took away from attending Drug Discovery Chemistry in 2024
Drug Discovery Chemistry is an annual event that focuses solely on early drug discovery, where scientists from around the world gather to showcase their research and participate in thoughtful...
NanoTemper Technologies introduced Monolith X, adding breakthrough Spectral Shift technology to their top-selling Monolith product line.
2:20Introducing Monolith X - the only way to get binding affinities in solution using very minimal sample
NanoTemper’s Monolith wins top global design award: Red Dot Award
NanoTemper’s newest Monolith instrument has been awarded the Red Dot Award, an internationally recognized seal of excellence in design. Both bold aesthetics and user-friendly design have always...
10 terms to help you more easily understand targeted protein degradation
If you’re new to the world of TPD or you’re trying to keep up with all developments in this fast-moving field, here are some terms and definitions that will help you.
14:01The desire to use technology that allows you to study membrane proteins in close-to-native conditions is specifically important when it comes to interaction measurements. It’s always a question how mu
35:47Targeted protein degradation using molecular glues or proteolysis-targeting chimeras (PROTACs) is an increasingly important therapeutic modality, especially for undruggable targets. Even with candidat
Why so many companies are betting on PROTACs as a superior therapeutic modality
One advantage of PROTACs is their unique mode of action — the induced proximity of an E3 ligase and the protein of interest (POI) triggers the ubiquitination and then degradation of the POI by the...
35:24Effect of different SARS-CoV-2 variants on COVID 19 assays and vaccines
SARS-CoV-2, the causative agent of COVID-19, has an ssRNA genome of almost 30kb that encodes for a total of 26 proteins. Among the four structural viral protein, in particular the S protein stands out
MST and TRIC technology to reliably study PROTAC binary and ternary binding in drug development
27:20Explore the complexities and drivers of molecular interactions with Dr. Tom Laue from the University of New Hampshire. He presented as a keynote speaker in the Protein Stability & Function: Can't have