Browse views: by Year, by Function, by GLF, by Subfunction, by Conference, by Journal

Affordable uniform isotope labeling with 2H, 13C and 15N in insect cells.

Sitarska, Agnieszka and Klopp, Julia and Roest, Susan and Skora, Lukasz and Fernandez, Cesar and Shrestha, Binesh and Gossert, Alvar (2015) Affordable uniform isotope labeling with 2H, 13C and 15N in insect cells. J Biomol NMR, 62 (2). pp. 191-197. ISSN 1573-5001

Abstract

A simple and affordable protocol was developed to produce uni-form labeled proteins in insect cells. Incorporation levels of 80% can be achieved for 15N and 13C and yields are comparable to expression in full media. For 2H,15N and 2H,13C,15N labeling, incorporation is slightly lower with 75% and 73%, respectively, and yields are typically two-fold reduced. The media were optimized for simplicity, reproducibility, isotope incorporation and cost. They consist of only five components, which are all commercially available at less than 8% of the costs than labeling media from vendors. High isotope incorporation and low cost are achieved by using labeled algal amino acid extracts and exploiting well-known biochemical pathways. The approach was applied to several cytosolic and secreted target proteins.

Item Type: Article
Date Deposited: 14 Jun 2016 23:45
Last Modified: 14 Jun 2016 23:45
URI: https://oak.novartis.com/id/eprint/25093

Search

Email Alerts

Register with OAK to receive email alerts for saved searches.