Proteins differ greatly in their size, composition, and three-dimensional structure, even though they basically share a common set of only 20 amino acid building blocks linked through a backbone of peptide bonds. This is possible because a single protein can consist of hundreds of individual amino acids and a large number of additional modifications are also carried out, such as phosphorylation or glycosylation, which offers almost infinite possibilities for variation. This variety is an essential requirement for proteins to fulfil the wide array of functions needed for sustaining biological life. Depending on their functions, they can be categorised in different subgroups. Enzymes, for example, are proteins which catalyse chemical reactions, while receptors and signal proteins facilitate signal transduction.
Protein assays in biologic and medicinal research range from simple protein quantification to complex interaction studies of protein binding. Microplate reader-based protein quantification methods are widely used to analyse and characterise protein samples. They are often only used to standardise other analysis results to the mass used. For the non-specific protein mass determination assays with absorbance readout, such as Bradford or Lowry assays, are employed to quantify total protein amounts. Immunological ligand binding assays, such as ELISAs, can be used to identify and quantify specific proteins in a sample. Apart from these, advanced antibody-based methods using BRET or TR-FRET readouts offer even higher sensitivity and allow users to work with minimal sample volumes, ideally suited for high-throughput approaches. Such techniques can even be employed to selectively detect specific protein modifications like for example protein phosphorylation.
Next to the quantification of proteins using microplate readers a wide range of assays for measuring enzyme or receptor activity are available which allow further characterisation of a protein sample. Generally, these activities are realised by the interaction of a target protein with another binding partners, like another protein, DNA, or other molecules. Microplate reader-based protein interaction assays enable the study of such events in miniature format. Dedicated features such as reagent injection and ultra-fast signal detection available on BMG LABTECH microplate readers can be used to analyse protein interactions with even greater detail and sensitivity. These possibilities are particularly important for the identification of new drugs and treatment options as dysregulated protein interactions are often involved in disorders and disease.
Search our resources section for information about specific applications, literature citations, videos, blog articles and many other publications. Many of the resources provided are associated with current and previous instrument models and versions.
Antibiotic resistance continues to increase. Find out here how BOCILLIN can be used to study resistant penicillin binding protein variants using fluorescence polarization.
Have you heard of dTAG degraders yet? Check out this novel approach in targeted protein degradation and learn how it can be used to identify disease-relevant proteins.
Receptor signalling is essential in physiological processes. Read here how a cell-based NanoBiT receptor dimerisation assay can be used to screen for androgen disruptors on the CLARIOstar Plus.
Read here how peptidic inhibitors of α-Synuclein nucleation and fibril elongation can be evaluated on microplate readers.
Looking for alternative acceptor fluorophores in TR-FRET attempts? Read here how CELT-335 was successfully used in a Cannabinoid competition binding assay at CB1.
Read here, how a fluorescence polarization-based interaction assay on the PHERAstar FSX can be used to reduce sample volumes, cost, and time-requirements for high-throughput screening.
Innovation for targeted protein degradation and next-generation degraders is gathering pace. This blog introduces some of the different approaches that act via the lysosome or proteasome.
The choice of assay for targeted protein degradation studies is crucial. But what is the preferred assay and detection technology for your specific research needs and microplate reader?
Molecular glues are small molecules that help target unwanted proteins for destruction by the ubiquitin-proteasome system. Find out how microplate readers can advance molecular glue research.
PROTACs are small, readily designed molecules that target unwanted proteins to the cell’s ubiquitin-proteasome system for degradation. Find out how microplate readers can advance PROTAC research.
The tau protein plays a role in many neurological diseases and disorders. Find out about neuronal toxicity induced by tau and how microplate readers can aid tau research.
G protein-coupled receptors are a gateway to many cellular responses. Find out how microplate readers can be used to support GPCR research.
Martin Schwalm discusses measuring PROTAC-induced degradation, ternary complex formation kinetics and stability, and shows a complete pipeline using live cells.
Binding affinity is an important parameter in drug discovery. Find out, how microplate readers can be used for binding affinity determination and the advantages they offer.
Which approaches can be used to quantify proteins? The most common absorbance-based assays that can be measured on a microplate reader are discussed in this talk.
Helen Harrison, Director of Screening at Amphista Therapeutics, discusses targeted protein degradation and the discovery of drugs in this area.
Dmitry Veprintsev & David Sykes talk about GPCR Binding and Signalling Studies and how the PHERAstar FSX can help them.
Maria Arruda from the University of Nottingham tests natural products from the Brazilian biodiversity for drug discovery purposes.
Laura Kilpatrick, Chloe Peach & Carl White from University of Nottingham talk about their work with the receptors VEGFR2 and CXCR4.
Steven Charlton from the University of Nottingham & Excellerate Biosciences talks about Binding Kinetics and how the PHERAstar FSX can help him.
Kevin Pfleger (University of Western Australia) talks about his experience with BRET-based ligand binding on BMG LABTECH Microplate readers.
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