Extraction of proteins from extensively calcified osseous tissue, such as cortical bone has been particularly
challenging for traditional methods of sample preparation. However, a comprehensive proteomic analysis
of bone is only possible when the total protein constituency is effectively isolated. The efficiency of sample
preparation is therefore a critical component of the analytical process.
Historically, extraction of protein from bone required prolonged acid demineralization over several days to
enable complete penetration of histochemical reagents to cellular components. Here we describe a method
for the extraction of protein from ostrich tibia, which was used as a model sample to develop an extraction
process that uses pressure cycling technology (PCT) and also which obvia testhe need for acid
demineralization prior to extraction. The ability to extract proteins from bone without prior demineralization
offers important advantages in efficient representative extraction of protein and significant time savings
during sample preparation.
利用壓力循環(huán)技術(shù)(PCT)對(duì)皮質(zhì)骨中分離出來的蛋白和多肽進(jìn)行電泳分析
從皮質(zhì)骨等充分鈣化的骨質(zhì)組織中提取蛋白的方法極大地挑戰(zhàn)了傳統(tǒng)的樣品制備方法。然而這種方法只有在總的目標(biāo)蛋白有效分離的時(shí)候,廣泛的蛋白質(zhì)組學(xué)分析才能成為可能。因此,樣品制備的有效性成為分析過程中的關(guān)鍵因素。歷*,從骨頭中提取蛋白需要幾天的去除礦物質(zhì)的過程使得組織化學(xué)試劑*滲透到 細(xì)胞組分中。本文描述了一種從鴕鳥脛骨中提取蛋白的方法,該方法利用了壓力循環(huán)技術(shù)(PCT),而且省去了以往的在提取蛋白之前先對(duì)樣品進(jìn)行去除礦物質(zhì)的 過程。這種方法使得提取蛋白非常有效而且節(jié)約了大量的時(shí)間成本。
Liberty研究級(jí)微波多肽合成儀
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