Příprava vzorku pro analýzu proteomu leukemických buněk BM2 dvourozměrnou elektroforézou
| Title in English | Sample preparation for proteom analysis of leukemic cells BM2 using two-dimensional electrophoresis |
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| Authors | |
| Year of publication | 2004 |
| Type | Article in Proceedings |
| Conference | 1. česká proteomická konference |
| MU Faculty or unit | |
| Citation | |
| Field | Oncology and hematology |
| Keywords | Two-dimensional electrophoresis; proteome analysis; sample preparation |
| Description | Quality of two-dimensional (2D) electrophoresis is dependent on the way of sample preparation. The aim of this study was to set up optimal conditions for preparation of the sample of v-myb-transformed chicken BM2 monoblasts to get a high number of resolved proteins in 2D gel. Starting composition of both lysis (9 M urea, 2 % NP-40, 3 % CHAPS, 70 mM DTT and 2 % ampholyte pH 8-10.5) and rehydratation buffers (8 M urea, 4 % CHAPS, 100 mM DTT, 0.001 % bromphenol blue and 0.2 % ampholyte pH 3-10) have been modified by addition of thiourea causing clear increase of the number of protein spots detectable in 2D gel. In addition, sonication of the sample further increased the number and density of resolved proteins. The presence of thiourea in the lysis/rehydratation buffers and sonication increased the number of protein spots resolved in 2D gel about 8-fold. These experimental conditions are going to be used in analysis of proteome during terminal differentiation of BM2 monoblasts. |
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