電極緩沖液中甘氨酸的作用

2017-05-11

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甘氨酸同Tris(三羥甲基氨基甲烷)一起組成電泳緩沖液中的緩沖對,可以穩(wěn)定電泳過程中的PH值。
在SDS-PAGE不連續(xù)電泳中,制膠緩沖液使用的是Tris-HCL緩沖系統(tǒng),濃縮膠是pH6.7,分離膠pH8.9;
而電泳緩沖液使用的Tris-甘氨酸緩沖系統(tǒng)。在濃縮膠中,其pH環(huán)境呈弱酸性,因此甘氨酸解離很少,其在電場的作用下,泳動(dòng)效率低;而CL離子卻很高,兩者之間形成導(dǎo)電性較低的區(qū)帶,蛋白分子就介于二者之間泳動(dòng)。由于導(dǎo)電性與電場強(qiáng)度成反比,這一區(qū)帶便形成了較高的電壓梯度,壓著蛋白質(zhì)分子聚集到一起,濃縮為一狹窄的區(qū)帶。當(dāng)樣品進(jìn)入分離膠后,由于膠中pH的增加,呈堿性,甘氨酸大量解離,泳動(dòng)速率增加,直接緊隨氯離子之后,同時(shí)由于分離膠孔徑的縮小,在電場的作用下,蛋白分子根據(jù)其固有的帶電性和分子大小進(jìn)行分離。

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