| Semi-thin Sectioning, Light and Fluorescence Microscopy of Floral Bud to Study Microspore Development in Arabidopsis
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Author:
Date:
2016-03-05
[Abstract] Pollen grains are male gametophytes produced within the pollen sacs of the anthers of the flower. Recent genetic studies have revealed several components involved in microspore development (Borg et al., 2009; Berger and Twell, 2011), and yet many components controlling microspore development remain to be identified. Semi-thin sectioning of anthers and light and fluorescence microscopy of floral bud (Kim et al., 2015) are the initial key experiments to characterize Arabidopsis mutants and transgenic plants for understanding the roles of new genetic components during microspore development. Herein, we describe a protocol for semi-thin sectioning of anthers and light and fluorescence microscopy of floral bud in Arabidopsis.
[摘要] 花粉粒是在花的花药的花粉囊内产生的雄性配子体。 最近的遗传研究揭示了涉及小孢子发育的几种组分（Borg等人，2009; Berger和Twell，2011），然而仍有许多控制小孢子发育的组分被鉴定。 花药的半薄切片和花芽的光和荧光显微镜检查（Kim等人，2015）是表征拟南芥突变体和转基因植物的初始关键实验 在小孢子发育过程中新遗传组分的作用。 在这里，我们描述了拟南芥花药半花瓣和光和荧光显微镜的半薄切片的协议。
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| In Gel Detection of Lipase Activity in Crude Plant Extracts (Olea europaea)
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Author:
Date:
2015-04-20
[Abstract] Here, we provide a detailed protocol describing a SDS-PAGE based procedure to assay in gel neutral lipase activity. Total protein extracts are separated by SDS-PAGE and gels are treated with lipase substrate-α-naphthyl palmitate. This long-chain fatty acid ester is hydrolysed by lipases present in the gel. The product resulting from this reaction can be then visualized in the gel as yellow-brownish activity bands. This relatively simple and effective method of lipase assay detection can be used for crude protein extracts from different plant tissues.
[摘要] 在这里，我们提供详细的协议描述基于SDS-PAGE的程序来测定凝胶中性脂肪酶活性。 通过SDS-PAGE分离总蛋白提取物，并用脂肪酶底物-α-棕榈酸萘酯处理凝胶。 这种长链脂肪酸酯被存在于凝胶中的脂肪酶水解。 由该反应产生的产物然后可以在凝胶中显现为黄棕色活性带。 这种相对简单和有效的脂肪酶测定法可用于来自不同植物组织的粗蛋白提取物。
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| Measurement of TACE Activity in Extracts from Cultured Cells
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Author:
Date:
2014-10-20
[Abstract] In cigarette smoke–induced and inflammation-associated lung cancer development, cigarette smoke extract (CSE) activates tumor necrosis factor-alpha (TNF-α) secretion from macrophages. TNF-α converting enzyme (TACE), also known as α-Secretase or ADAM17 (A Disintegrin and Metalloprotease), is a member of the ADAM family of metalloproteases. TACE mediated ectodomain shedding leads to the conversion of the inactive TNF-α precursor into the active mature pro-inflammatory cytokine. The SensoLyte 520 TACE (α-Secretase) Activity Assay Kit was used to detect TACE activity in CSE-activated macrophages. This assay is reliable, reproducible and easy to carry out in 96 well plate format.
[摘要] 在香烟烟雾诱导和炎症相关的肺癌发展中，香烟烟雾提取物（CSE）激活巨噬细胞的肿瘤坏死因子-α（TNF-α）分泌。 TNF-α转化酶（TACE），也称为α-分泌酶或ADAM17（解聚素和金属蛋白酶），是金属蛋白酶的ADAM家族的成员。 TACE介导的胞外结构域脱落导致无活性的TNF-α前体转化为活性成熟促炎细胞因子。 SensoLyte 520 TACE（α-分泌酶）活性测定试剂盒用于检测CSE活化的巨噬细胞中的TACE活性。 该测定法是可靠的，可重复的并且容易在96孔板形式中进行。
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