Laboratórna Diagnostika
Published December 6, 2021 | Version v1
Journal article Open

Optimalizácia metodík SDS-PAGE a želatínovej zymografie na rýchlu proteomickú analýzu pacientov / Optimalization of SDS-PAGE and gelatin zymography methods for rapid proteomic analysis of patients

Authors/Creators

  • 1. Ústav lekárskej a klinickej biochémie, UPJŠ LF, Košice

Description

SÚHRN

Cieľom predkladanej práce bolo vyvinúť účinný protokol polyakrylamidovej gélovej elektroforézy na báze dodecylsulfátu sodného (SDS-PAGE), ako aj želatínovej zymografie, na detekciu proteínov. Na štúdium elektroforetického profilovania proteínov bolo použitých 100 vzoriek od pacientov s kolorektálnym karcinómom (CRC) v rôznych štádiách. Bol navrhnutý presný protokol na optimalizáciu podmienok, ako je pH používaných roztokov, koncentrácie jednotlivých zložiek gélov, množstvo vzorky nanášanej do gélu, časové rozmedzie farbenia, odfarbovania a iné. Optimalizáciou týchto faktorov boli metódou zymografie detegované matrixové metaloproteinázy v svojej latentnej aj aktívnej forme.

ABSTRACT

            The present work aimed to develop an efficient protocol for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), as well as gelatin zymography, for the detection of proteins. To study the electrophoretic profiling of proteins, 100 samples from patients with colorectal carcinoma (CRC) at different stages were used. An accurate protocol has been proposed to optimize conditions such as the correct pH level of the solutions used, the concentration of the individual components of the gels, the amount of sample applied to the gel, the time range of staining, decolorization, etc. By optimizing these factors, matrix metalloproteinases in both latent and active forms were detected by zymography.

Notes

Článok bol realizovaný s podporou grantového projektu VEGA 1/0333/20. The article was implemented with the support of the grant project VEGA 1/0333/2

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