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Aphidicolin from Nigrospora sphaerica

来自黑孢菌的Aphidicolin

公司名称: Sigma-Aldrich
产品编号: A0781
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Immunostaining of Formaldehyde-fixed Metaphase Chromosome from Untreated and Aphidicolin-treated DT40 Cells
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Date:
2017-05-05
[Abstract]  During mitosis chromosomes are condensed into dense X-shaped structures that allow for microscopic determination of karyotype as well as inspection of chromosome morphology.

This protocol describes a method to perform immunostaining of formaldehyde-fixed metaphase chromosomes from the avian cell line DT40. It was developed to characterize the localization of YFP-tagged TopBP1 on mitotic chromosomes and specifically determine the percentage of TopBP1 foci that formed on breaks/gaps as well as ends of individual metaphase macrochromosomes (Pedersen et al., 2015). For this ...
[摘要]  在有丝分裂期间,染色体被浓缩成致密X型结构,允许显微检测染色体核型和检查染色体形态。
该方案描述了从禽细胞系DT40进行甲醛固定的中期染色体的免疫染色的方法。 它被开发用于表征YFP标记的TopBP1在有丝分裂染色体上的定位,并且具体确定形成于断裂/间隙以及单个中期大染色体末端的TopBP1灶的百分比(Pedersen等,2015)。 为此,应用YFP的免疫染色。 然而,该协议可以针对其他细胞系或表位进行优化。
【背景】染色中期染色体的显微镜分析是经典的细胞遗传学技术,广泛用于研究和诊断。基本原理包括通过锭子不稳定试剂如谷氨酸诱导中期细胞周期停滞,这将触发主轴装配检查点,从而阻止细胞在中期。这用于富集浓缩染色体的细胞。随后将细胞在低渗溶液中进行溶胀,然后在显微镜载玻片上铺展有丝分裂细胞。最终结果是来自单细胞的显微镜检测染色体,便于染色体核型分析以及个体染色体的研究。传统上,肿胀的细胞用甲醇和乙酸(3:1)固定,然后铺展在载玻片上(Hungerford,1965; Ronne等,1979)。
这里描述的方法使用甲醛而不是甲醇进行固定。这可以用于随后用与甲醇固定不相容的抗体染色。该方案针对鸡DT40细胞的中期传播进行了优化,YFP标记的TopBP1在中期大染色体上的免疫染色(Pedersen等,2015)。 ...

Synchronize Human Embryonic Stem Cells at Different Cell Cycle Stage
Author:
Date:
2012-06-05
[Abstract]  Pluripotency and the capability for self-renewal are essential characteristics of human embryonic stem cells (hESCs), which hold great potential as a cellular source for tissue replacement. Short cell cycle (15-16 h) compared to somatic cells is another property of hESCs. Efficient synchronization of hESCs at different cell cycle stages is important to elucidate the mechanistic link between cell cycle regulation and cell fate decision. This protocol describes how to establish synchronization of hESCs at different cell cycle stages. [摘要]  人胚胎干细胞(hESCs)的基本特征是多潜能性和有自我更新的能力,有潜力作为组织替换的细胞供源。和体细胞相比,hESCs有很短的细胞周期这是它的另一个主要体征。不同hESCs细胞周期的高效的同步化对于阐明细胞周期调控和细胞分化之间的关系机制十分的重要。

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