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970603
授課教師 :
海洋生物技術系
蔡志明
Electrophoretic methods
Simple electrophoresis describes the
situation in which the protein
molecules are separated in an
electric field at a constant pH and
current
簡單的描述電泳,就是在電場利用固定的 pH
值和電流使蛋白質分子分離。
Isoelectric focusing 等電聚焦
Electrophoretic steps are not often
employed in protein purification 電泳
步驟在蛋白質純化不是經常利用的。
Analysis for purity
Electrophoretic analysis 電流生理學
Resolution of separate components of a protein
mixture is achieved most clearly using an
electrophoresis method
利用電泳方法,可以清楚得到蛋白質的組成分析。
Preparative electrophoresis has problems in the
design and use of apparatus, so despite its
obvious potential, it is not often used
儘管電泳方法非常有用,但麻煩在機器的設計與用途,因
此不常使用。
But in an analytical mode, electrophoresis is a
most widely method 但是在分析的方法中,電泳是最
普遍的。
It is almost obligatory to characterize a purified
protein preparation by an electrophoretic
technique
它的特性是利用電泳技術來純化蛋白質。
Gel system 凝膠系統
Before gel system were developed,
electrophoretic analysis was carried out in the
Tiselius free-boundary apparatus, requiring tens
of milligrams of proteins 在發展凝膠系統前,電泳分
析經過 Tiselius free-boundary 儀器,需要數十毫克的
蛋白質 。
Starch gel 澱粉凝膠
Synthetic gel medium- crossed linked
polyacrylamide 合成凝膠 - 經過聚丙烯
酰胺連結
Polyacrylamide gel electrophoresis
聚丙烯酰胺凝膠電泳
Simple (native) gel electrophoreis
簡單的凝膠電泳
聚丙烯酰胺凝膠電泳在十二烷基硫酸鈉
Although separation is only on the
basis of size, resolution of closely
similar components is excellent 雖然
分離只能用在基本的大小規模,但是分析
類似成分的能力卻是優秀的。
The charge/size ratio is virtually
identically for all proteins, and
separation can occur only as a result
of the molecular sieving through the
pores of the gel 所有蛋白質的電荷與大
小比例幾乎是相同的,分離作用會在分子
篩選通過凝膠的孔隙時發生。
Gradient gel
Native protein 天然蛋白質
It is run until no further movement occurs,
and a “focusing” or concentration of
diffuse protein zones can occur, resulting
in very fine resolution 直到沒有一個“重點”
或散開的蛋白區濃度可以持續運轉時,會產生了
極微細的分析。
Gradient-SDS gel 有梯度的 SDS 凝膠
Isoelectric focusing
Isoelectric focusing involves setting
up a pH gradient and allowing the
proteins to migrate in an electric field
to the point in the system where the
pH equals their isoelectric point
在 pH 值等於其等電點時,等電聚焦成立一
個 pH 梯度,並允許蛋白質遷移在一電場點
。
Immobilized pH gradient, IPG
固定化 pH 梯度
Two-dimensional systems
兩維系統
Staining and detection of proteins after
electrophoresis
染色和檢測的蛋白質電泳