The major protein expression profile and two‐dimensional protein databse of human heart

Abstract

The construction of a two-dimensional protein database of the human heart is presented. The database contains information on about 300 abundant proteins of human myocardial tissue, including approximately 40 proteins that were identified by different methods. Each protein was characterized according to several parameters, including molecular weight, isoelectric point, name, partial sequence, subcellular localization, and genetic as well as embryonic changes.

Study of Dj-1 protein in tissue specimens, cultured cells and serum of prostate cancer patients

“…Приготовление белковых экстрактов, их фракционирование методом двумерного электрофореза по О'Фарреллу, визуализацию белков окрашиванием кумасси голубым R-250 и азотнокислым серебром и анализ полученных двумерных электрофореграмм (ДЭ) выполняли как описано ранее [13,14]. При исследовании каждого биоптата было получено не менее трёх ДЭ, а при изучении белков клеточных линий -не менее 10 для каждой.…”

“…При исследовании каждого биоптата было получено не менее трёх ДЭ, а при изучении белков клеточных линий -не менее 10 для каждой. Идентификацию белков методами MALDI-TOF MS и MS/MS масс-спектрометрии проводили на MALDI-времяпролетном масс-спектрометре Ultraflex ("Bruker", Германия) с УФ-лазером (336 нм) в режиме положительных ионов в диапазоне масс 500-8000 Да с калибровкой их по известным пикам аутолиза трипсина [13,14]. Для определения молекулярных масс белковых фракций использовали наборы высокоочищенных рекомбинантных белков: "SM0661", (10-200 кДа); "SM0671", (10-170 кДа); ("Fermentas", США).…”

Identification of Proteins Involved in pH Adaptation in Extremophile Yeast Yarrowia lipolytica

“…A well was created for protein marker application at the edge of each gel slab. Further details of the modified 2DE approach are described earlier (Kovalyova et al, 1994;Laptev et al, 1995;Kovalyov et al, 1995). For protein visualization, the polyacrylamide gel slabs were stained with Coomassie Blue R-250 and then with silver nitrate according to the well-described methods (Blum et al, 1987) and modified by the addition of 0.8% acetic acid to sodium thiosulfate.…”

Comparative Proteomic Study of Proteins in Prostate Cancer and Benign Hyperplasia Cells

“…Another variant (IF-2DE) was a special modification of O'Farrell method which used nonequilibrium pH gradient electrophoresis (NEPHGE) as was described earlier [18]. In this variant IEF was performed in glass tubes (2.4 × 180 mm) filled with 4% PAAG prepared on 9М urea solution containing 2% of triton Х-100 and 2% mixture of ampholines.…”

“…Protein visualization by Coomassie Blue R-250 and sodium nitrate staining as well as analysis of two-dimensional (2D) electrophoregrams were performed as described earlier [17,18] with minor modifications. At least three 2D electrophoregrams were obtained for each biopsy specimen, whereas the study of cultured cells proteins was performed using at least ten 2D electrophoregrams.…”