Knowing the strength of the interactions between key players will give you the insights you need to understand the details behind how biological systems work.
If you’re having difficulty studying challenging interactions that include membrane proteins, PROTACs, intrinsically disordered proteins (IDPs), or RNA-based therapeutics, you’ll need Monolith to characterize them.
Monolith measures the broadest range of binding affinities in-solution, using very little sample
Measure high and low affinities in the same instrument
Different projects have different demands. It’s inevitable that you’ll need to look at both weak and strong molecular interactions. With a broad sensitivity range, Monolith measures Kds varying from pM to mM, so you can tackle as many projects as you want.
Characterize binding events with very little sample
It’s not easy to prepare the large volumes of highly concentrated target and ligand required by ITC. With Monolith, you get a Kd in a fraction of the volume and concentration, leaving you with more sample for additional experiments.
Measure in solution, in close-to-native conditions
Immobilizing molecules in SPR assays often causes them to lose activity, and you’re left without results. Because Monolith measures in solution in most buffers, both binding partners are free to interact in their native conformation, you’ll finally get results.
Characterize interactions with many different types of molecules or samples
You know what types of interactions you need to characterize now, but it’s always difficult to predict what you’ll need to look at in the future. Monolith gives you one less thing to worry about because it has the flexibility to handle all different types of molecules and samples.
Membrane proteins, intrinsically disordered proteins (IDPs), receptors, enzymes, antibodies, and nanobodies
Fragments, PROTACs, ions, nanoparticles, peptides, and carbohydrates
DNA, RNA, and aptamers
Exosomes and liposomes
Platelets and whole cells
Virus particles and empty capsids
Evaluate more than a binary interaction
Assess relative affinities of two or more molecules for the same target
Ternary binding events
Characterize interactions that involve three or more binding partners
Derive additional information from your affinity assays
Oligomerization and aggregation
Monitor these events to understand protein functionality
Calculate molecular ratios of binding partners
Derive ∆G, ∆H and ∆S from calculated Kds
*Requires offline data handling, not supported by Monolith software
Use MST technology to measure interactions
Monolith uses MST technology to quantify molecular interactions between a target and ligand by detecting changes in fluorescence intensity while a temperature gradient is applied over time (grey box, top figure). The fluorescent signal comes from the target that is either fluorescently labeled or has intrinsic fluorescence and becomes an extremely sensitive reporter for the interaction. The binding affinity is automatically determined at the end of each run without additional and lengthy data analysis. The affinity constant (Kd) is calculated from a fitted curve that plots normalized fluorescence against concentration of ligand (bottom figure).
Monolith has no fluidics — that means there’s really no regular maintenance
Life is so much easier when fluidics aren’t involved. Monolith doesn’t require cleaning or flushing in between runs, or a maintenance contract. So, it’s ready whenever you’re ready.
The best choice for studying interactions spanning a broad range of sensitivity. Turn pico mode on to measure very strong interactions, or off for weak ones.
Choose this if you want the most flexibility when it comes to labeling strategies. Pick the two fluorescent channels you need the most.
When you want to measure interactions by detecting the intrinsic fluorescence of your target protein, choose this.
As interactions are characterized free in solution, MST has been successful where SPR failed due to the limitations of a surface-based approach.
Dr. Stephan Uebel
Max-Planck-Institute of Biochemistry, Germany
Feel confident your experiments will run smoothly with software that’s smart
Most software starts once you load your samples and start your run. Monolith’s MO.Control is built differently — not only do you get help with guided step-by-step experimental planning and assay setup before you start your run, but you also get immediate feedback on assay optimization based on your results after the run is over. MO.Control 2 adds the ability to optimize buffer conditions more efficiently so you can get to generating results faster.
MO.Affinity Analysis 3
Make sure your analysis is consistent across various data sets and that you’re identifying any insights across replicate measurements. MO.Affinity Analysis 3 is the ideal complement to MO.Control 2 — merge and group your data sets for comparison purposes and then easily report results with presentation-worthy data and publication-ready figures.
Get great results with tailor-made consumables
Monolith capillaries are made with care — they’re manufactured in a state-of-the-art facility and rigorously tested. Pair the capillaries with one of the Monolith Protein Labeling Kits to get the highest quality data and ultimately, the best outcome with Monolith.
Spend less time optimizing your assay and more time getting actionable results
Spending time and money just optimizing assays and not generating usable data is super-frustrating, especially when you have limited resources. Monolith has ways to help you start with the target labeling step, and with optimizing your assays conditions so you can get to actionable results sooner.
Your success starts with choosing the right labeling strategy
The online and interactive Protein Labeling Assistant gives you the best labeling recommendations for your target protein and your instrument optics.
Feel confident you have the right buffer conditions
MO.Control 2 software makes it super easy to check the effects of buffers on your interactions. Take advantage of the ability to run 24 capillaries to screen up to 6 different buffer conditions in duplicates and with no-ligand controls in the same run. Read the technical note for an example of how easy it is to find the right buffer conditions.
Find the optimal buffer conditions in less time
Use the Buffer Exploration Kit, a 96-well plate loaded with ready-to-use buffers commonly used in MST assays so you don’t have to pre-mix them yourself. It makes buffer screening less time-consuming.
Get instant quality checks
Key quality parameters such as aggregation, photobleaching, and low signal-to-noise are constantly monitored for you. If additional optimization is necessary, you’ll get recommendations on how to correct these issues.