| Western Blot Analysis of Chloroplast HSP70B in Chlorella Species
|
|
Author:
Date:
2013-08-05
[Abstract] Western blotting allows for the specific detection of proteins by an antibody of interest. This protocol utilizes isolation of total proteins protocol for Chlorella vulgaris prior to gel electrophoresis. After electrophoresis, the selected antibodies are used to detect and quantify levels of chloroplast HSP70B.
[摘要] 蛋白质印迹允许目标抗体对蛋白质的特异性检测。 该方案在凝胶电泳之前利用用于小球藻的总蛋白质方案的分离。 电泳后，所选择的抗体用于检测和定量叶绿体HSP70B的水平。
|
|
|
| Generation of Polyclonal Specific Antibodies
|
|
Author:
Date:
2013-06-05
[Abstract] Generation of antibodies specific for a protein of interest is a common method in many disciplines. This protocol details the steps in production of a polyclonal antibody in rabbits using a bacterially expressed fusion protein as an antigen. The protocol is generated based on data presented in Wirschell et al.(2013).
[摘要] 对感兴趣的蛋白质特异性的抗体的产生是许多学科中的常见方法。 该方案详述了使用细菌表达的融合蛋白作为抗原在兔中产生多克隆抗体的步骤。 该协议基于Wirschell等人（2013）中提供的数据生成。
|
|
|
| GTP Binding Assays in Arabidopsis thaliana
|
|
Author:
Date:
2013-05-05
[Abstract] Signaling through G proteins constitutes an ancient mechanism that functions in the transduction of extracellular signals into intracellular responses. Activation of a proper receptor by a stimuli leads to an exchange of GDP for GTP, the activation of G proteins and the dissociation of Gα-GTP from the Gβγ dimer for the heterotrimeric G proteins. The G protein subunits remain active until the intrinsic GTPase activity or/and a GTPase activating protein result in the hydrolysis of GTP to GDP and the inactivation of the protein. Here we describe a protocol for measuring GTP binding activity from Arabidopsis plant protein extracts using GTP γ35S. GTP γ35S assay measures the level of G protein activation following a stimuli, by determining the ...
[摘要] 通过G蛋白的信号传导构成了在将细胞外信号转导到细胞内反应中起作用的古老机制。通过刺激激活适当的受体导致GTP的GDP交换，G蛋白的激活和G emα-GTP从G emβγ二聚体的解离异源三聚体G蛋白。 G蛋白亚基保持活性，直到内在的GTP酶活性或/和GTP酶活化蛋白导致GTP水解为GDP和蛋白质的失活。在这里，我们描述了使用GTPγ S，从拟南芥植物蛋白提取物测量GTP结合活性的方案。 GTPγ测定通过测定不可水解的类似物GTPγ35 S的结合活性来测量刺激后G蛋白活化的水平。为了确定特异性G蛋白活性，应包括特异性突变体和/或过表达体提取物，并作为对照测量。
|
|
|