Cell Meter 荧光法细胞周期检测试剂盒 405nm激

---

流式细胞仪
Ex:	405 nm
Em：	450/40 nm
通道:	Pacific Blue 通道

样品实验方案

简要概述

1. 用5×10⁵至1×10⁶细胞/ mL的密度制备含测试化合物的细胞
2. 在0.5 mL细胞溶液中加入1 µL 500X Nuclear Violet
3. 在室温下孵育30-60分钟
4. 使用405nm紫色光激发流式细胞仪进行分析

 

实验步骤

1.对于每个样品，请在0.5 mL的温暖培养基或自备缓冲液中以5×10⁵至1×10⁶细胞/ mL的密度制备细胞。注意：应单独评估每种细胞系，以确定诱导凋亡的细胞密度。

2.用测试化合物处理细胞，以诱导凋亡或其他细胞周期功能。

3.向含有生长培养基的细胞中加入1µL 500X Nuclear Violet （组分A），并在37°C，5％CO₂的培养箱中孵育细胞30至60分钟。注意：对于贴壁细胞，用0.5mM EDTA轻轻提起细胞以保持细胞完整，并在用Nuclear Violet 孵育之前用含血清的培养基洗涤一次。注意：适当的孵育时间取决于所用的单个细胞类型和细胞浓度。优化每个实验的孵育时间。注意：DNA染色前无需固定细胞，因为Nuclear Violet 具有细胞渗透性。

4.可选：以1000 rpm离心细胞4分钟，然后将细胞重悬于0.5 mL检测缓冲液（组分B）或自备缓冲液中。

5.使用405 nm紫色激光（Ex / Em = 405/445 nm）通过流式细胞仪检测荧光强度。

 

参考文献

Cell cycle synchronization of Escherichia coli using the stringent response, with fluorescence labeling assays for DNA content and replication
Authors: Ferullo DJ, Cooper DL, Moore HR, Lovett ST.
Journal: Methods (2009): 8

DNA replication, cell cycle progression and the targeted gene repair reaction
Authors: Engstrom JU, Kmiec EB.
Journal: Cell Cycle (2008): 1402

Morin inhibits the growth of human leukemia HL-60 cells via cell cycle arrest and induction of apoptosis through mitochondria dependent pathway
Authors: Kuo HM, Chang LS, Lin YL, Lu HF, Yang JS, Lee JH, Chung JG.
Journal: Anticancer Res (2007): 395

Direct control of cell cycle gene expression by proto-oncogene product ACTR, and its autoregulation underlies its transforming activity
Authors: Louie MC, Revenko AS, Zou JX, Yao J, Chen HW.
Journal: Mol Cell Biol (2006): 3810

Cell cycle markers for live cell analyses
Authors: Easwaran HP, Leonhardt H, Cardoso MC.
Journal: Cell Cycle (2005): 453

Dynamic relocalization of hOGG1 during the cell cycle is disrupted in cells harbouring the hOGG1-Cys326 polymorphic variant
Authors: Luna L, Rolseth V, Hildrestr and GA, Otterlei M, Dantzer F, Bjoras M, Seeberg E.
Journal: Nucleic Acids Res (2005): 1813

Dynamics of relative chromosome position during the cell cycle
Authors: Essers J, van Cappellen WA, Theil AF, van Drunen E, Jaspers NG, Hoeijmakers JH, Wyman C, Vermeulen W, Kanaar R.
Journal: Mol Biol Cell (2005): 769

Cell cycle regulation of the murine 8-oxoguanine DNA glycosylase (mOGG1): mOGG1 associates with microtubules during interphase and mitosis
Authors: Conlon KA, Zharkov DO, Berrios M.
Journal: DNA Repair (Amst) (2004): 1601

Description of a flow cytometry approach based on SYBR-14 staining for the assessment of DNA content, cell cycle analysis, and sorting of living normal and neoplastic cells
Authors: Nunez R, Garay N, Villafane C, Bruno A, Lindgren V.
Journal: Exp Mol Pathol (2004): 29

Differential roles of STAT1alpha and STAT1beta in fludarabine-induced cell cycle arrest and apoptosis in human B cells
Authors: Baran-Marszak F, Feuillard J, Najjar I, Le Clorennec C, Bechet JM, Dusanter-Fourt I, Bornkamm GW, Raphael M, Fagard R.
Journal: Blood (2004): 2475