Scientists choose Monolith as their primary tool to measure binding interactions

Measure the broadest range of binding affinities in solution.

Get Kds in minutes using very little sample.

Evaluate the most difficult targets
Small molecules, GPCRs, even targets like whole viral particles and cells

Measure any sample type
From cell cultures, to fungi, plant extracts and bioliquids

Consume very little sample
Don’t use up all your precious sample in one assay. Get a Kd with just a tiny amount

Reduce time spent in assay development
Work with the buffer that keeps your protein stable even if that means adding DMSO

Get results quickly
Capturing measurements in minutes means you can make better decisions on what to do next, sooner

Find out which ligands bind more tightly
Sensitivity down to pM gives you confidence when measuring strong affinities

Measure in close to native conditions
No immobilization is required

Measure even the weakest interactions
Evaluate mM-level affinities for ligands like fragments

 


Always be ready for your next project

Because Monolith is free of fluidics, there is no cleaning, priming, calibration or flushing needed between runs or even between projects.

 

Monolith NT.115

Most versatile
Crowd favorite for binding interactions

Monolith NT.115Pico

High sensitivity
Detect affinities in low pM range

Monolith NT.LabelFree

Dye-free
Analyze label-free using intrinsic fluorescence

Monolith NT.Automated

Workhorse
Fast screening with reliable results. Add the NT.Automated Screening Unit for fully automated sample and liquid handling

The Monolith NT.115 system is easy to use and only takes a short amount of time to set up experiments. The sample consumption is very low and its immobilization-free approach makes handling protein samples and setting up assays less complicated. The technical support and customer service from NanoTemper are excellent. Overall, we are very satisfied and strongly recommend the system to research groups looking for a fast quantitative binding assay.

Professor Antonio Macchiarulo
Department of Pharmaceutical Sciences, University of Perugia, Italy

 

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All it takes is 10 minutes per Kd

Monolith is all about determining binding affinity at the end of each run without additional and lengthy data analysis. The strength of the interaction between a fluorescently labeled or intrinsically fluorescent sample and a binding partner (or ligand) is measured by detecting changes in the fluorescence intensity while a temperature gradient is applied over time (grey box, left figure). From this, binding affinity (Kd) is automatically calculated from a fitted curve that plots normalized fluorescence against concentration of ligand (right figure).

 

Monolith delivers more than just a Kd

Competition assays
Assess relative affinities of two or more molecules for the same target

Single-dose and affinity screens
Find hits and rank them based on their Kd

Oligomerization and folding
Monitor these events to understand protein functionality

Ternary binding events
Characterize interactions that involve three or more binding partners

Stoichiometry*
Calculate molecular ratios of binding partners

Thermodynamics*
Derive ∆G, ∆H and ∆S from calculated Kds

*Requires offline data handling, not supported by Monolith software

Understand how molecular complexes interact

Dr. Dong and colleagues from the Harbin Institute of Technology wanted to understand how LbCpf1 (a Cas-9-like nuclease) recognizes crRNA. After solving the crystal structure of the crRNA-Cpf1 complex, they looked at the role Mg2+ played in stabilizing the crRNA conformation and its interaction with Cpf1, and like many other scientists they chose Monolith to measure the binding affinity. The interaction between crRNA and LbCpf1 was reduced ~50-fold as compared to that in the absence of EDTA, showing that the ion was crucial for the stabilization of the crRNA-endonuclease complex.

Monitoring MG2+ dependence of CRISPR RNA to a Cas9-like nuclease
Dong et. al., Nature, 532:522-536 2016

Measure interactions on whole cells

Like other scientists, Prof. Lindahl and Dr. Macwan at Linköping University want to understand how the molecules involved in biological processes interact with each other. They used Monolith to measure the affinity between CD42b – a receptor expressed on the surface of human platelets – and labeled anti-CD42b antibody. Monolith enabled them to measure this interaction directly on the surface of whole platelets, bringing them one step closer on their quest to understanding hemostasis and thrombosis.

Monolith MST cell interactions

Data obtained in collaboration with Prof. T. Lindahl and Dr. A. Macwan. Department of Clinical and Experimental Medicine, Linköping University.

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Accuracy is what counts

Monolith uses MicroScale Thermophoresis (MST) technology to quantify binding events. One partner is typically labeled with a fluorescent dye and becomes an extremely sensitive reporter for all binding interactions. These events, weak or strong, cause a change in MST signal that is easily detected. The results are automatically translated into precise and quantitative Kd values.

Getting these accurate results quickly requires the use of high quality capillaries. Advantages are numerous—minute amounts of samples are analyzed in-solution, under native or close-to-native conditions, and in an immobilization-free environment without the need for purification.

 

Software that makes sense

All Monolith systems, no matter the throughput, come with control software that makes sense. For those large data sets from screening experiments, MO.Screening Analysis software just might be the right thing to add to your tool box.

 


MO.Control

MO.Control software makes it so anyone can get Kds. With guided step-by-step planning and assay setup, users feel confident their experiments will run smoothly. Get binding values instantly and one-click exportable results.
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MO.Affinity Analysis

As the ideal complement to MO.Control, MO.Affinity Analysis enables advanced data analysis like merging, grouping, or comparing data sets. Easily report results with presentation-worthy data and publication-ready figures.

MO.Screening Control

Get similar step-by-step guidance that made MO.Control popular. MO.Screening Control, designed specifically for screening experiments, helps with planning sample layout on either 96- or 384-well plates, defining measurement settings and performing measurements.

MO.Screening Analysis

Let MO.Screening Analysis automatically evaluate and identify the hits among thousands of ligands from single dose screens. Identify strong and weak ligands by reporting Kds in affinity-based screenings.
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Great results with tailor-made consumables

Monolith capillaries are made with care—they’re made in a state-of-the-art facility and rigorously tested. Pair them with one of the Monolith Protein Labeling Kits and any of the Monolith systems to get the highest quality data and ultimately, the best outcome.

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