Fluorescence-Based Assays for Kinase Activity

Fluorescence-Based Assays for Kinase Activity

INQUIRY

Creative BioMart offers high-quality services on kinase activity studies using florescence-based assays to assist our customers in their kinase biology research and drug discovery. Our talented scientific team is able to provide our customers with unparalleled expertise and experience in kinase biology and pharmaceutical research.

Background

Kinases, which constitute one of the most abundant gene families in humans, are one of the largest families of evolutionarily related proteins. The human kinome consists of over 500 different protein kinases and lipid kinases. In addition to a conserved ATP binding site, each kinase has one or more substrate-binding site(s). Kinases can alter the functional activity of proteins in many ways and are involved in the regulation of many cellular events in eukaryotic cells, such as cell cycle progression, transcription, metabolism, and apoptosis. There has been much clinical evidence that kinases are the most effective therapeutic targets in various types of diseases. Due to the large size of the kinase family, multiple techniques to study kinase activity have been extensively studied and developed. However, a large number of different assay formats poses a challenge in deciding which one will be the best fit.

Radioisotope assays using radioactive 32P and immunoassays using phosphate-specific antibodies are conventional techniques for effective detection of kinase activity; however, their complex labeling procedures may increase assay time and associated costs, and the use of radioisotopes can pose a threat to human health. Therefore, efforts have been made to develop sensitive, simple, and cost-effective assays. Fluorescence offers unmatched sensitivity, stability, and flexibility compared to many other techniques. The diversity of available fluorescent dyes provides virtually unlimited possibilities for assay design. These advantages make fluorescence-based assays one of the easiest and popular methods for kinase activity studies.

Featured Services

Creative BioMart offers various types of fluorescence-based assays for kinase activity to meet the specific needs of our customers. Our scientists will work with you to develop an optimum assay. If you can’t find an assay of your interest, please fill out the online inquiry form and let us know more details. We can offer you our expertise to help you accelerate your success.

Assay Types	Details
Fluorescent Peptide Assays	Monitoring the incorporation of phosphate into a peptide substrate. A class of fluorescent peptide substrate as alternative of frequently used radioisotopes.
Molecular Beacon Probes	Molecular beacons (MBs) with high sensitivity and excellent specificity. A simple, fast, and sensitive assay using MBs to detect protein kinase activity.
Fluorescence Intensity Assay (FI)	Commonly used fluorescence-based methods for detecting kinase activity. A homogenous assay to produce a simple fluorescent intensity output.
Fluorescence Polarization (FP) Assay	Monitoring a fluorescent molecular movement and rotation. Widely used in kinase inhibitor screening.
Fluorescent Polymer Superquenching	Enhanced sensitivity and biological specificity are provided by monitoring fluorescence changes. A homogeneous and robust kinase assay for use in multi-well plate format.
Fluorescence Lifetime Technology (FLT)	An alternative to traditional fluorescence-based readouts. A robust and economical platform for kinase screening and profiling.
Fluorescence Resonance Energy Transfer (FRET)	Kinase reactions can be quantified by calculating the ratio of donor emission to acceptor emission after excitation of the donor fluorophore. Homogenous format and simple application to HTS.
Time-Resolved Fluorescence (TRF) Assay	Fluorophores such as lanthanide ions with long fluorescence decay time are used. Sensitive and free of interference from molecules with a very different fluorescence lifetime.
Time-Resolved Fluorescence Resonance Energy Transfer (TR-FRET)	The practical combination of TRF with FRET. A peptide substrate labeled with an acceptor fluorophore and an anti-phospho-peptide detection antibody labeled with a donor fluorophore are used.
Graphene Oxide-Based Multiplexed Fluorescence Assay	A novel graphene oxide sensing platform for the multiplexed detection of kinases. Developed as a routine tool for the high-throughput screening for kinases inhibitors.

Creative BioMart is an advanced biotech company within the field of kinase/phosphatase biology. We offer our expertise in establishing assays for kinase screening & profiling. We have state-of-the-art facilities and technology to provide efficient services with guaranteed turnaround times. A large collection of ready-to-use kinase activity assay kits are also available to help our customers save precious time. If you have any questions, please do not hesitate to contact us.

References

• Wang L, et al. A label-free and sensitive fluorescent assay for one step detection of protein kinase activity and inhibition. Analytica chimica acta, 2016, 935: 224-230.
• Wang Y & Ma H. Protein kinase profiling assays: a technology review. Drug Discovery Today: Technologies, 2015, 18: 1-8.