Our website does not fully support your browser.

We've detected that you are using an older version of Internet Explorer. Your commerce experience may be limited. Please update your browser to Internet Explorer 11 or above.

Promega's Cookie Policy

We use cookies and similar technologies to make our website work, run analytics, improve our website, and show you personalized content and advertising. Some of these cookies are essential for our website to work. For others, we won’t set them unless you accept them. To find out more about cookies and how to manage cookies, read our Cookie Policy.

Improved In-Gel Digestion Results and Work-Flow Through the Use of a Mass Spectrometry Compatible Surfactant

Part # PS094

Abstract

Daniel J. Simpson1, Sergei Saveliev1, Becky Godat1, and Grzegorz Sabat2
1Promega Corporation, Madison, WI
2University of Wisconsin, Madison, WI

Typical protocols for in-gel protein digestion require overnight incubation followed by additional 2–3 hours for peptide extraction. We present here a protocol and reagent to streamline this process and in many cases improve protein coverage. The reagent, ProteaseMAX™ Surfactant, enhances peptide cleavage by providing a denaturing environment prior to protease addition. It is also designed to degrade during the proteolytic reaction, generating species that do not interfere with mass spectrometry. For in-gel applications, complete digestions can be performed in 1 hour and the surfactant provides concurrent peptide extraction. This obviates the requirement for separate extraction processing, greatly simplifying the overall work-flow. Recovery is particularly enhanced for longer and more hydrophobic peptides (2,500–4,000Da), thereby improving coverage.

Printed in USA.