Abstract
INTRODUCTIONActivation and inactivation of proteins using photoactivation of caged peptides or proteins offer insights into cellular dynamics not achievable using genetic means. The ability to selectively alter the activity of a specific protein at a defined time and location inside a cell allows the correlation of changes in protein activity and cellular behavior. A caged compound, peptide, or protein is prepared by covalently linking it to a photolabile, protecting group via a limited number of critical functional groups in the biomolecules. Under carefully defined conditions, a brief irradiation with UV light (320-400 nm) can lead to an intramolecular photoisomerization reaction that induces the cleavage of the chemical bond between the cage group and the unmodified, biologically active species. The following protocol uses a semiautomated microinjection system (Eppendorf FemtoJet Microinjector along with the Eppendorf InjectMan) for the introduction of caged compounds into cells. Unlike the bead-loading approach, this technique allows the investigator to choose which cells are targeted.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.