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Glycerol, AR® (ACS)

Company: Avantor Performance Materials
Catalog#: 5092
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Modification of 3’ Terminal Ends of DNA and RNA Using DNA Polymerase θ Terminal Transferase Activity
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Date:
2017-06-20
[Abstract]  DNA polymerase θ (Polθ) is a promiscuous enzyme that is essential for the error-prone DNA double-strand break (DSB) repair pathway called alternative end-joining (alt-EJ). During this form of DSB repair, Polθ performs terminal transferase activity at the 3’ termini of resected DSBs via templated and non-templated nucleotide addition cycles. Since human Polθ is able to modify the 3’ terminal ends of both DNA and RNA with a wide array of large and diverse ribonucleotide and deoxyribonucleotide analogs, its terminal transferase activity is more useful for biotechnology applications than terminal deoxynucleotidyl transferase (TdT). Here, we present in detail simple methods by which purified human Polθ is utilized to modify the 3’ terminal ends of RNA and DNA for various applications in ... [摘要]  DNA聚合酶θ(Polθ)是一种混杂的酶,对易错的DNA双链断裂(DSB)修复途径而言是必需的,称为替代性末端连接(alt-EJ)。 在这种形式的DSB修复中,Polθ通过模板和非模板核苷酸添加循环在切割的DSB的3'末端处进行末端转移酶活性。 由于人Polθ能够用广泛的多种核糖核苷酸和脱氧核糖核苷酸类似物修饰DNA和RNA的3'末端,因此其末端转移酶活性对于生物技术应用比末端脱氧核苷酸转移酶(TdT)更有用。 在这里,我们详细介绍使用纯化的人Polθ修饰生物技术和生物医学研究中各种应用的RNA和DNA的3'末端的简单方法。
【背景】人类POLQ基因编码含有N末端超家族2(SF2)型解旋酶结构域和C末端A家族聚合酶结构域的大蛋白质(Sfeir和Symington,2015; Black et al。,2016; Wood and Doublie, 2016)。蛋白质也编码一个大的中心结构域,其功能尚未归入。 Polθ在后生动物中表达,已被证明在DNA复制和修复的多个方面起作用(Black et al。,2016; Wood and Doublie,2016)。最近的工作表明,哺乳动物Polθ对于易于识别的DNA双链断裂(DSB)修复途径而言是必需的,称为替代性末端连接(alt-EJ),也称微小鼠介导的终结合(MMEJ)(Yousefzadeh et al。 ,2014; ...

Muscle Histology Characterization Using H&E Staining and Muscle Fiber Type Classification Using Immunofluorescence Staining
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Date:
2017-05-20
[Abstract]  Muscle function is determined by its structure and fiber type composition. Here we describe a protocol to examine muscle histology and myofiber types using hematoxylin and eosin (H&E) and immunofluorescence staining, respectively. H&E stain nucleus in blue and cytoplasm in red, therefore allowing for morphological analyses, such as myofiber diameter, the presence of degenerated and regenerated myofibers, and adipocytes and fibrotic cells. Muscle fibers in adult skeletal muscles of rodents are classified into 4 subtypes based on the expression of myosin heavy chain proteins: Myh7 (type I fiber), Myh2 (type IIA fiber), Myh1 (type IIX fiber), Myh4 (type IIB fiber). A panel of monoclonal antibodies can be used to specifically label these muscle fiber subtypes. These protocols are commonly ... [摘要]  肌肉功能由其结构和纤维类型组成决定。 这里我们描述一个协议,分别检查使用苏木精和伊红(H&E)和免疫荧光染色的肌肉组织学和肌纤维类型。 H&E在蓝色和细胞质红色染色核,因此允许形态学分析,如肌纤维直径,退化和再生肌纤维的存在,以及脂肪细胞和纤维化细胞。 基于肌球蛋白重链蛋白Myh7(I型纤维),Myh2(IIA型纤维),Myh1(IIX型纤维),Myh4(IIB型纤维)的表达,将啮齿动物成年骨骼肌肌肉纤维分为4种亚型:。 一组单克隆抗体可用于特异性标记这些肌纤维亚型。 这些方案通常用于肌肉发育,生长和再生的研究(例如:Wang等,2015; Nie等,2016; Yue et al。,2016; Wang et al。,2017)。
【背景】骨骼肌由肌细胞,脂肪细胞,成纤维细胞和其他细胞类型组成。 多核心肌细胞,骨骼肌的主要组成,也称为肌纤维(肌纤维)。 肌肉组织学研究是肌肉功能研究的常规方法。 一般来说,肌纤维的直径和脂肪细胞的程度和纤维化面积与肌肉力量有关(Yue et al。,2016)。 此外,中心有核肌纤维的存在作为肌肉再生期间新再生肌纤维的替代指标(Wang等,2017)。 基于差异代谢性状和肌球蛋白重链(MyHC)亚型的表达,肌纤维分为四种类型(I,IIa,IIx和IIb)。 ...

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