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.

Rapid Quantitation of IgG after Digestion at Elevated Temperature with a Novel Trypsin Reagent

Part # PS298

Abstract

Michael M. Rosenblatt1, Sergei Saveliev1, Daniel Spellman2, Kevin Bateman2, and Marjeta Urh1
1Promega Corporation, 2800 Woods Hollow Rd, Madison, WI, 53711
2Merck & Co., Incorporated , West Point PA, 19486

As increasing numbers of protein-based therapeutics enter development pipelines, more efficient protocols are required for structural characterization and quantitation. Proteolysis of these proteins into peptides represents a bottleneck and often requires optimization of numerous steps including reduction, alkylation and digestion time. We have developed a new Trypsin reagent, Rapid Digestion--Trypsin and Rapid Digestion--Trypsin with Lys C, that streamlines the entire sample preparation process to less than 1 hour. With this new process, proteolysis is performed at 70°C which facilitates both denaturation and rapid digestion. This protocol is robust, amenable to multiple analytes including pure proteins and complex mixtures, and is compatible with digestion under native, reduced and/or denaturing conditions.

Printed in USA.