Abstract
The aim of this study is to develop a novel method for detection of glycoproteins on polyacrylamide gel. In this method, radio-iodinated-tyrosine (125I-tyrosine) was conjugated to glycoprotein by schiff's base mechanism on the sodium dodecyl sulfate- polyacrylamide gel. Ovalbumin and Concanavalin A (Con A) were used as a glycosylated and a non-glycosylated model proteins, respectively. The proteins were separated in SDS- PAGE and oligosaccharides on the glycoprotein were oxidised using periodic acid to produce aldehydes than 125I-tyroine was conjugated to aldehyde groups without using reducing agent like Sodium Metabisulfite. The radio-iodinated glycoprotein on gel was scanned using a Multi-Photon Detection (MPD) scanner. The elechtrophoretic analysis of ovalbumin and Con A were performed and stained with Coomassie brilliant blue to identify total proteins, while MPD detection of glycoproteins using 125I-tyrosine selectively detected ovalbumin. Present results showed that MPD enhanced glycoprotein detection method can be used as a sensitive tool for the detection of glycoproteins on polyacrylamide gel
| Original language | English |
|---|---|
| Journal | International Journal of Biological Chemistry |
| Volume | 3 |
| Issue number | 1 |
| Pages (from-to) | 18-24 |
| Number of pages | 7 |
| ISSN | 1819-155X |
| Publication status | Published - 2009 |