𝔖 Bobbio Scriptorium
✦   LIBER   ✦

In-gel deglycosylation of sodiumdodecyl sulfate polyacrylamide gel electrophoresis-separated glycoproteins for carbohydrate estimation by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

✍ Scribed by S. Kilz; H. Budzikiewicz; S. Waffenschmidt

Publisher
John Wiley and Sons
Year
2002
Tongue
English
Weight
209 KB
Volume
37
Category
Article
ISSN
1076-5174

No coin nor oath required. For personal study only.

✦ Synopsis

Abstract

Mass determination by mass spectrometric methods (electrospray ionization mass spectrometry (ESI‐MS), matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOFMS)) of sodiumdodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE)‐separated proteins is a well known procedure and reliable protocols are available. In our efforts to use the established methods to determine the molecular mass of the disulfide brigded, heterodimeric glycoprotein GP3 and to determine the carbohydrate content of each protein subunit we developed an in‐gel chemical deglycosylation method. For this purpose we established experimental conditions that allow maximum extraction of the high molecular mass protein subunits and developed a routine method to apply the HF–pyridine deglycosylation protocol to proteins isolated from polyacrylamide gel pieces. The novel protocol and extraction procedure described can be used to analyze O‐glycosylated proteins up to 150 kDa after SDS‐PAGE separation. Copyright © 2002 John Wiley & Sons, Ltd.

📜 SIMILAR VOLUMES

Identification of Staphylococcus aureus
Identification of Staphylococcus aureus exotoxins by combined sodium dodecyl sulfate gel electrophoresis and matrix-assisted laser desorption/ ionization-time of flight mass spectrometry
✍ Katussevani Bernardo; Silke Fleer; Norbert Pakulat; Oleg Krut; Frank Hünger; Mar 📂 Article 📅 2002 🏛 John Wiley and Sons 🌐 English ⚖ 313 KB 👁 2 views

Staphylococcus aureus is an important human pathogen whose pathogenesis involves the synthesis of cell wall associated virulence factors and secreted toxins with damaging effects on the host cells. Most of these pathogenic factors are synthesized in a growth-phase dependent manner as a response to e

Electroelution and passive elution of γ-
Electroelution and passive elution of γ-globulins from sodium dodecyl sulphate polyacrylamide gel electrophoresis gels for matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry
✍ Marina Galvani; Mahmoud Hamdan 📂 Article 📅 2000 🏛 John Wiley and Sons 🌐 English ⚖ 130 KB 👁 1 views

To the Editor-in-Chief Sir, Electroelution and passive elution of g-globulins from sodium dodecyl sulphate polyacrylamide gel electrophoresis gels for matrix-assisted laser desorption/ionisation time-offlight mass spectrometry Figure 1. (a) SDS-PAGE stained with Coomassie Brilliant Blue of intact hu

Large-gel two-dimensional electrophoresi
Large-gel two-dimensional electrophoresis-matrix assisted laser desorption/ionization-time of flight-mass spectrometry: An analytical challenge for studying complex protein mixtures
✍ Eckhard Nordhoff; Volker Egelhofer; Patrick Giavalisco; Holger Eickhoff; Martin 📂 Article 📅 2001 🏛 John Wiley and Sons 🌐 English ⚖ 400 KB 👁 2 views

## Large-gel two-dimensional electrophoresis-matrix assisted laser desorption/ionization-time of flight-mass spectrometry: An analytical challenge for studying complex protein mixtures The large-gel two-dimensional electrophoresis (2-DE) technique, developed by Klose and co-workers over the past 2