Dulbecco’s Modified Eagle’s Medium - high glucose

Dulbecco''s Modified Eagle''s Medium - 高葡萄糖

Company: Sigma-Aldrich

Catalog#: D0422

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Monitoring Protein Splicing Using In-gel Fluorescence Immediately Following SDS-PAGE

Joel Weinberger II and Christopher W. Lennon

Published online: 2021-08-20

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Monitoring the Targeting of Cathepsin D to the Lysosome by Metabolic Labeling and Pulse-chase Analysis

Author:

Lucas A. Tavares and Luis L. P. daSilva,

Date:

2017-11-05

[Abstract] Mannose 6-phosphate receptors function can be studied in living cells by investigating alterations in processing and secretion of their ligand Cathepsin D. The assay described here is well established in the literature and comprises the metabolic labeling of newly synthesized proteins with [³⁵S] methionine-cysteine in HeLa cells to monitor Cathepsin D processing through secretory pathway and secretion using immunoprecipitation, SDS-PAGE and fluorography. [摘要] 通过研究其配体组织蛋白酶D的加工和分泌的改变，可以在活细胞中研究甘露糖-6-磷酸受体功能。在此描述的测定在文献中已经很好地确定，并且包括新合成的蛋白质的代谢标记，其中[35 [S]甲硫氨酸半胱氨酸，以监测组织蛋白酶D处理，通过分泌途径和分泌使用免疫沉淀，SDS-PAGE和荧光。

【背景】组织蛋白酶d（CATD）是一种溶酶体天冬氨酸蛋白酶由甘露糖-6-磷酸受体（M6PRs）排序在哺乳动物细胞中该传输它从反面高尔基体网络到内涵体/溶酶体（戈什等人，2003 ）。将CatD合成为前体蛋白（〜52kDa），其在溶酶体中被切割以产生中间体（〜48kDa）或成熟的溶酶体形式（〜34kDa）。微量的前体蛋白质也从生物合成途径分泌（Benes et al。，2008）。 catD的丰度可以使用几种方法来确定，例如基于免疫荧光的染色（Poole等人，1972），蛋白质印迹，荧光活性测定（Bewley等人，（Hirst等人，2009; Kametaka等人，2007; Tavares等人，2011）或用脉冲追踪分析进行代谢标记（Hirst等人，2009; Kametaka等人， ...

Protein Translation Study – Label Protein with S35 Methionine in Cells

Author:

Salma Hasan and Isabelle Plo,

Date:

2012-11-05

[Abstract] To follow protein synthesis, cells should be incubated with radioactive amino acid such as [35S] methionine during mRNA translation. Then, the neosynthetized protein will be identified by an autoradiography after immunoprecipitation with a specific antibody and separation on a polyacrylamide denaturing gel. [摘要] 为了遵循蛋白质合成，在mRNA翻译期间，细胞应与放射性氨基酸如[35S]甲硫氨酸一起温育。然后，在用特异性抗体免疫沉淀并在聚丙烯酰胺变性凝胶上分离后，通过放射自显影鉴定新合成的蛋白质。

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Dulbecco’s Modified Eagle’s Medium - high glucose

Dulbecco''s Modified Eagle''s Medium - 高葡萄糖

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