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PENICILLIN-STREPTOMYCIN SOLUTIO

青霉素/链霉素溶液

Company: WISENT
Catalog#: 450-201-EL
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Uptake Assays to Monitor Anthracyclines Entry into Mammalian Cells
Author:
Date:
2017-09-20
[Abstract]  Anthracyclines, such as doxorubicin and daunorubicin, are DNA damaging agents that autofluoresce and can be readily detected in cells. Herein, we developed suitable assays to quantify and localize daunorubicin in mammalian cells. These assays can be exploited to identify components that are involved in the uptake of anthracyclines. [摘要]  蒽环类药物,如多柔比星和柔红霉素,是DNA损伤剂,其可自发荧光并且可以容易地在细胞中检测到。 在这里,我们开发了合适的测定法来定量和定位哺乳动物细胞中的柔红霉素。 可以利用这些测定来鉴定参与蒽环类药物吸收的成分。
【背景】蒽环类药物,如多柔比星和柔红霉素,通过损伤DNA起作用,用于治疗各种类型的癌症,包括急性骨髓性白血病。当癌症患者被系统地给予蒽环类抗生素时,有几个因素限制了到达肿瘤部位的药物的量(Chauncey,2001; Deng等,2014; Riganti等,2015)。在肿瘤中,对癌细胞的药物活性受到药物吸收不良,药物流出过量以及细胞靶标变化的限制。几十年来,这些DNA损伤剂如何进入癌细胞还不清楚(Aouida等,2010; Cesar-Razquin等,2015; Zhang等,2015)。为了解决这个问题,我们开发了三种可靠的体外测定法来监测柔红霉素在细胞中的积累。这些测定中的两个是定量的,需要获得荧光活化细胞分选(FACS)口径和Fluoroskan仪器,第三个是使用落射荧光显微镜进行半定量。使用这些测定法,我们确定柔红霉素以时间和浓度依赖的方式进入细胞,并且每种细胞类型显示不同的摄取速率,这表明活性过程参与蒽环类药物的摄入(Andreev等人, ...

Analysis of Enteric Neural Crest Cell Migration Using Heterotopic Grafts of Embryonic Guts
Author:
Date:
2016-09-05
[Abstract]  Hirschsprung disease (HSCR), also named aganglionic megacolon, is a severe congenital malformation characterized by a lack of enteric nervous system (ENS) in the terminal regions of the bowel (Bergeron et al., 2013). As the ENS notably regulates motility in the whole gastrointestinal track, the segment without neurons remains tonically contracted, resulting in functional intestinal obstruction and accumulation of fecal material (megacolon). HSCR occurs when enteric neural progenitors of vagal neural crest origin fail to fully colonize the developing intestines. These “enteric” neural crest cells (ENCCs) have to migrate in a rostro-caudal direction during a fixed temporal window, which is between embryonic day (e) 9.5 and e14.5 in the mouse (Obermayr et al., 2013). ... [摘要]  Hirschsprung病(HSCR),也称为神经节巨结肠,是严重的先天性畸形,其特征在于在肠的末端区域缺乏肠神经系统(ENS)(Bergeron等人,2013)。由于ENS特别调节整个胃肠道的运动性,没有神经元的节段保持紧张收缩,导致功能性肠梗阻和粪便材料(巨结肠)的积累。 HSCR发生在迷走神经嵴来源的肠神经祖细胞不能完全殖民发育的肠道。这些"肠"神经嵴细胞(ENCC)必须在固定的时间窗口内沿着尾 - 尾方向迁移,其在小鼠的胚胎天(e)9.5和e14.5之间(Obermayr等, ,2013)。最近,我们的小组产生了称为荷斯坦的新的HSCR小鼠模型,其中ENCC的迁移受损,因为它们的微环境中胶原VI水平增加(Soret等人,2015)。在这里,我们描述了允许我们演示这种迁移缺陷的细胞自主性的方法。在从先前描述的异位移植方法改造的该系统中(Breau等人,2006),供体组织是e12.5中肠的完全定殖的区段,而宿主组织是e12的非耳部分.5后肠。在宿主组织中ENCC迁移的程度在培养24小时后评估,并且当供体组织具有转基因背景例如允许内源标记的Gata4-RFP(Pilon等人,2008)时极大地促进ENCC在宿主组织中的迁移的程度的ENCCs。根据供体和宿主组织的遗传背景,这种方法可以允许评估细胞自主和非细胞自主缺陷的ENCC迁移。

Cell-based Assays to Monitor AID Activity
Author:
Date:
2016-02-05
[Abstract]  The enzyme Activation induced deaminase (AID) underpins antibody affinity maturation and isotype switching through its mutagenic activity of deaminating deoxycytidine to deoxyuridine in DNA. Subsequent processing of the deoxyuridine initiates the processes of somatic hypermutation (SHM) and class switch recombination (CSR) in B cells. Structure-function analysis of AID requires sensitive and biologically relevant methods to measure its various activities. Here we describe simple but effective methods to measure 1) the ability of AID to mutate the Escherichia coli genome, which provides an indication of its catalytic activity; 2) the capacity of AID to perform SHM by complementing a derivative of the DT40 chicken B cell line; 3) the ability of AID to perform CSR by complementing ... [摘要]  酶活化诱导的脱氨酶(AID)通过其将脱氧胞苷脱氨基到DNA中的脱氧尿苷的诱变活性来支持抗体亲和力成熟和同种型转换。脱氧尿苷的后续加工引发B细胞中体细胞超突变(SHM)和类型转换重组(CSR)的过程。 AID的结构功能分析需要灵敏和生物相关的方法来测量其各种活动。在这里我们描述简单但有效的方法来测量1)AID突变大肠杆菌基因组的能力,其提供其催化活性的指示; 2)AID通过补充DT40鸡B细胞系的衍生物来进行SHM的能力; 3)AID通过补充AID缺乏的原代小鼠B细胞来进行CSR的能力。三种方法的组合,伴随着AID亚细胞定位和蛋白质表达水平和稳定性的必要分析作为对照,允许AID的详细结构功能研究。

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