2D Diagonal Redox SDS-PAGE of Proteins
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Author:
Date:
2013-06-05
[Abstract] 2D diagonal redox SDS-PAGE of proteins is used to detect intramolecular or intermolecular disulfide bridges using Chlamydomonas in this example (Stroeher and Dietz, 2008; Schwarz et al., 2012). Both dimensions consist of a conventional SDS-PAGE, except that the sample buffer for the first dimension lacks a reducing agent. Intermolecular disulfide bridges increase the apparent molecular weight of a protein in the first dimension, whereas intramolecular bridges decrease the apparent weight of the protein.
[摘要] 在该实施例中,使用蛋白质的2D对角线氧化还原SDS-PAGE用于检测分子内或分子间二硫键(Stroeher和Dietz,2008; Schwarz等人,2012) 。 两个维度由常规SDS-PAGE组成,除了第一维的样品缓冲液不含还原剂。 分子间二硫桥在第一维中增加蛋白质的表观分子量,而分子内桥降低蛋白质的表观重量。
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35S pulse Labelling of Chlamydomonas Chloroplast Proteins
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Author:
Date:
2013-06-05
[Abstract] 35S pulse labelling of proteins is used to attach a radioactive label to newly synthesized proteins, as sulfur is an element that is mainly present in proteins (Fleischmann and Rochaix 1999). Depending on your organism’s uptake mechanisms you need cysteine, methionine or sulfuric acid as a source of radioactive sulfur. This example uses Chlamydomonas cells and H235SO4 (Schwarz et al., 2012).
[摘要] 由于硫是主要存在于蛋白质中的元素(Fleischmann和Rochaix 1999),因此蛋白质的 S脉冲标记用于将放射性标记附着到新合成的蛋白质上。 根据你的生物体的吸收机制,你需要半胱氨酸,甲硫氨酸或硫酸作为放射性硫的来源。 该实施例使用衣原体细胞和H 2 35 SO Schwarz等人 。,2012)。
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