| α2β1-integrin Clustering and Internalization Protocol
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Author:
Date:
2014-04-05
[Abstract] α2β1-integrin clustering experiment can be used to trigger internalization of α2β1-integrin. When clustering is performed with sequential administration of primary and fluorescent secondary antibodies, the entry kinetics of integrin can be followed into the cell. The idea is first to allow binding of primary antibodies (recognizing the extracellular epitope) to the α2β1-integrins and then to cluster the α2β1-integrin-bound primary antibodies together by the means of the secondary antibody. Binding is done on ice so that the α2β1-integrins will not internalize before both sets of antibodies are bound. Clustering is known to trigger α2β1-integrin internalization efficiently from the cell surface to the cytoplasm. In this protocol we used antibody-induced clustering of α2β1-integrin in order ...
[摘要] α2β1整合素聚类实验可用于触发α2β1整联蛋白的内化。 当通过顺序施用原发性和荧光二抗进行聚类时，可以在细胞中进入整联蛋白的进入动力学。 该想法是首先允许一级抗体（识别胞外表位）与α2β1整联蛋白的结合，然后通过二级抗体将α2β1整联蛋白结合的一级抗体聚集在一起。 在冰上进行结合，使得α2β1-整联蛋白在两组抗体结合之前不内化。 聚类已知能够有效地从细胞表面向细胞质触发α2β1整合素内化。 在这个协议中，我们使用抗体诱导聚类的α2β1整合素，以定量内化的α2β1整合素的数量与细胞表面相关的α2β1整合素相比。
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| Catalase Activity Assay in Candida glabrata
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Author:
Date:
2014-03-20
[Abstract] Commensal and pathogenic fungi are exposed to hydrogen peroxide (H2O2) produced by macrophages of the host. Pathogenic fungi counteract the harmful effects of H2O2 with the enzyme catalase (EC 1.11.1.6), which decomposes two molecules of H2O2 to two molecules of H2O and O2. Contribution of antioxidant systems on fungal virulence is actively studied. Measurement of catalase activity can contribute to the elucidation of the factors that influence the regulation of this pivotal enzyme. Here we describe a simple spectrophotometric method in which the activity of catalase is measured in total yeast extracts. Decomposition of H2O2 by the yeast extract is followed by the decrease in ...
[摘要] 共生和致病真菌暴露于由宿主的巨噬细胞产生的过氧化氢（H 2 O 2 O 2）。 致病真菌抵消了H 2 O 2对于过氧化氢酶（EC 1.11.1.6）的有害影响，所述过氧化氢酶分解两个分子的H 2 O 2 - O 2至两个H 2 O和O 2分子。 积极研究抗氧化系统对真菌毒力的贡献。 过氧化氢酶活性的测量可有助于阐明影响这种关键酶的调节的因素。 在这里我们描述一个简单的分光光度法，其中过氧化氢酶的活性在总酵母提取物中测量。 通过酵母提取物的H 2 O 2 O 2分解后，在240nm处的吸光度降低。 吸光度随时间的差异（ΔA240）被推断为过氧化氢酶活性的量度。
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