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Gewählte Publikation:

Schriebl, K; Trummer, E; Weik, R; Müller, D; Kunert, R; Lattenmayer, C; Katinger, H; Vorauer-Uhl, K.
(2006): A novel strategy for quantitative isoform detection directly performed from culture supernatant.
J Pharm Biomed Anal. 2006; 42(3):322-327 FullText FullText_BOKU

Abstract:
Currently, one of the most used techniques for the determination of isoform pattern analysis is isoelectric focusing. Routinely, this is performed by immunoblotting. Blotting of proteins after isoelectric focusing on IPG gels may cause several problems, such as protein loss by the blotting itself and band broadening, in some cases the immunostaining with antibodies might be problematic. In the present study, an alternative isoform prestaining method with CyDye fluors is presented. For this approach, a highly glycosylated fusion protein, Epo-Fc, was used consisting of two recombinant human erythropoietin attached to the Fc part of a human IgG(1) molecule. By using CyDye fluors, up to three samples can be focused on the same lane under identical electrophoretic conditions. A fundamental benefit of this technique is the ability to perform quantitative isoform pattern analysis directly from serum-free culture supernatant. (c) 2006 Elsevier B.V. All rights reserved.
Autor*innen der BOKU Wien:
Katinger Hermann
Kunert Renate
Trummer-Gödl Evelyn
Vorauer-Uhl Karola
BOKU Gendermonitor:

Find related publications in this database (using NML MeSH Indexing)
Animals -
CHO Cells -
Cricetinae -
Erythropoietin, Recombinant - analysis
Immunoglobulin Fc Fragments - analysis
Isoelectric Focusing - analysis
Protein Isoforms - analysis
Recombinant Fusion Proteins - analysis
Reproducibility of Results - analysis

Find related publications in this database (Keywords)
fusion protein Epo-Fc
CHO cells
isoelectric focusing
isoform pattern
CyDye fluors


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