NHS-ester-protein-labeling
Liv Jensen
Disclaimer
DISCLAIMER – FOR INFORMATIONAL PURPOSES ONLY; USE AT YOUR OWN RISK
The protocol content here is for informational purposes only and does not constitute legal, medical, clinical, or safety advice, or otherwise; content added to protocols.io is not peer reviewed and may not have undergone a formal approval of any kind. Information presented in this protocol should not substitute for independent professional judgment, advice, diagnosis, or treatment. Any action you take or refrain from taking using or relying upon the information presented here is strictly at your own risk. You agree that neither the Company nor any of the authors, contributors, administrators, or anyone else associated with protocols.io, can be held responsible for your use of the information contained in or linked to this protocol or any of our Sites/Apps and Services.
Abstract
Protocol for labeling a purified protein with an NHS ester fluorescent dye.
Steps
Mix 40μM unlabeled protein with 80μM ATTO 565 NHS ester dye in labeling buffer.
Incubate 1 hr at room temperature.
Buffer exchange the reaction into quench buffer over a pre-equilibrated G-25 desalting
column.
Assess labeling efficiency by measuring the ratio of absorbance at 280 and 564 nm,
correcting for dye absorbance at 280nm, using a Nanodrop spectrophotometer
