Protein phosphorylation, particularly on serine, threonine, or tyrosine residues, is a critical post-translational modification that serves as a major mechanism for regulating protein function. This modification plays a vital role in controlling various cellular processes such as apoptosis and signal transduction pathways. While dysregulated protein phosphorylation has been linked to the development of complicated diseases. Therefore, it is urgent to understand more details about phosphorylation.
Fig. 1 Typical functions of protein phosphorylation.1
To deeply understand protein phosphorylation, Creative Biolabs develops an effective in vitro cellular phosphorylation assay service for global customers.
Our in vitro cellular phosphorylation assay service is a cutting-edge cell-based kinase activity assay that enables the direct measurement of compounds' ability to inhibit kinase activity within a cellular environment. Our assay is capable of assessing phosphorylation using endogenously expressed target kinases or cells engineered for overexpression of the kinase under study. At the endpoint, we employ ELISA assay to quantify the substrate phosphorylation levels. By leveraging our in vitro cellular phosphorylation assay, customers are able to accurately test and screen potential therapeutics. Additionally, we offer a variety of kinase cell lines to choose from for your specific projects. At Creative Biolabs, our team of scientific experts is dedicated to delivering exceptional results tailored to our customers' specific needs and requirements.
Fig. 2 Working process of our in vitro cellular phosphorylation assay.
We deliver numerous potential targets to apply for diverse pathways research, which offers more convenience for global customers' project design.
Q1: What method do you use to induce protein phosphorylation in a laboratory setting?
A1: In vitro phosphorylation of proteins can be achieved by incubating the protein-of-interest with a specific kinase enzyme and ATP, leading to the addition of phosphate groups to specific residues on the protein. Negative controls with unphosphorylated versions of the protein need to be included for comparison.
Q2: How to measure phosphorylation in an in vitro setting?
A2: In vitro assay for phosphorylation utilizes purified proteins, isolated kinases, substrate peptides, and cofactors like ATP and Mg2+ ions. This approach enables detailed analysis of phosphorylation kinetics, stoichiometry, and target sequences. For more physiologically relevant assessments, isolated cells or tissues can be employed with appropriate controls to validate kinase activation.
For more details about our in vitro cellular phosphorylation assay service, please feel free to reach out to us.
References
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