|
||
| Journals and Publications |
|
AbstractElectrophoretic Characterisation of Hevein H.Y. Yeang, S.A.M. Arif, Y.H. Loke, N.P. Chew and S.M. Mohsin Although the latex protein, hevein, has received much research interest, its analysis by conventional SDS-polyacrylamide gel electrophoresis has been problematic. Hevein does not migrate at a rate that commensurates with its molecular weight and it does not always produce a sharp band on a polyacrylamide gel. To compound this problem, hevein is difficult to stain because it has a tendency to diffuse out of the gel during the de-staining process. Moreover, hevein Western blot is difficult to perform because, as a small protein, it easily passes through a nitrocellulose membrane during electro-transfer. Using the tricine SDS-polyacrylamide gel electrophoresis system that is optimised for small proteins and peptides, hevein migrated in the gel at a rate consistent with its molecular weight of 4.7 kDa, although the migration rate was significantly reduced under non-reducing conditions. Addition of phosphotungstic acid to the Coomassie blue stain fixed hevein and prevented its loss by diffusion out of the gel. Hevein could be effectively Western blotted on to the PVDF membrane and fixed with glutaraldehyde, after which, the protein was detectable using anti-hevein antibody. Although hevein is an unglycosylated protein, it was found to be associated with carbohydrate, probably through its lectin property in binding carbohydrate. |
|||
|
The Government and Malaysian Rubber Board shall not be liable for any loss or damage caused by the usage of any information obtained from this site. Malaysian Rubber Board 2002. All Rights Reserved. Best viewed with Internet Explorer 6.x browser.
|
||||