样品实验方案
工作溶液配制
TWO-PRO -3 工作溶液
在含20 mM Hepes缓冲的Hanks缓冲液(HH缓冲液)或您选择的其他缓冲液中,制备TWO-PROTM-3的工作液。
注意:在初始实验中,最好尝试几种染料浓度以确定产生结果的最佳浓度。高染料浓度往往会导致其他细胞结构的非特异性染色。
操作步骤
注意:以下实验步骤可用于大多数细胞类型。生长基质、细胞密度、其他细胞类型及生长因子的存在可能会影响染色效果。玻璃容器上残留的洗涤剂也可能影响许多组织的染色,并会使溶液中含有或不含细胞都出现强烈荧光物质。
1.根据需要培养和处理细胞。
2.除去细胞培养基。
3.将 TWO-PRO-3 工作溶液(1 至 10 µM)添加到细胞(悬浮细胞或贴壁细胞)中,并将细胞染色 15 至 60 分钟。
4.除去染料工作溶液并添加 HH 缓冲液或您选择的缓冲液。
5.使用 Cy5 滤光片通过荧光显微镜分析细胞染色。
试剂应用文献
Synergistic Activity and Mechanism of Sanguinarine with Polymyxin B against Gram-Negative Bacterial Infections
Authors: Qiao, Luyao and Zhang, Yu and Chen, Ying and Chi, Xiangyin and Ding, Jinwen and Zhang, Hongjuan and Han, Yanxing and Zhang, Bo and Jiang, Jiandong and Lin, Yuan
Journal: Pharmaceutics (2024): 70
A lysosomal regulatory circuit essential for the development and function of microglia
Authors: Iyer, Harini and Shen, Kimberle and Meireles, Ana M and Talbot, William S
Journal: Science advances (2022): eabp8321
参考文献
Daptomycin exerts rapid bactericidal activity against Bacillus anthracis without disrupting membrane integrity
Authors: Xing YH, Wang W, Dai SQ, Liu TY, Tan JJ, Qu GL, Li YX, Ling Y, Liu G, Fu XQ, Chen HP.
Journal: Acta Pharmacol Sin (2014): 211
Quantification of Candida albicans by flow cytometry using TO-PRO((R))-3 iodide as a single-stain viability dye
Authors: Kerstens M, Boulet G, Pintelon I, Hellings M, Voeten L, Delputte P, Maes L, Cos P.
Journal: J Microbiol Methods (2013): 189
A silicon cell cycle in a bacterial model of calcium phosphate mineralogenesis
Authors: Linton KM, Tapping CR, Adams DG, Carter RD, Shore RC, Aaron JE.
Journal: Micron (2013): 419
Pulsed electromagnetic field affects intrinsic and endoplasmatic reticulum apoptosis induction pathways in MonoMac6 cell line culture
Authors: Kaszuba-Zwoinska J, Chorobik P, Juszczak K, Zaraska W, Thor PJ.
Journal: J Physiol Pharmacol (2012): 537
Determination of the drug-DNA binding modes using fluorescence-based assays
Authors: Williams AK, Dasilva SC, Bhatta A, Rawal B, Liu M, Korobkova EA.
Journal: Anal Biochem (2012): 66
Transient changes in neuronal cell membrane permeability after blast exposure
Authors: Arun P, Abu-Taleb R, Valiyaveettil M, Wang Y, Long JB, Nambiar MP.
Journal: Neuroreport (2012): 342
A comparison of TO-PRO-1 iodide and 5-CFDA-AM staining methods for assessing viability of planktonic algae with epifluorescence microscopy
Authors: Gorokhova E, Mattsson L, Sundstrom AM.
Journal: J Microbiol Methods (2012): 216
SOCS-3 antagonizes pro-apoptotic effects of TRAIL and resveratrol in prostate cancer cells
Authors: Horndasch M, Culig Z.
Journal: Prostate (2011): 1357
The fluorescent dyes TO-PRO-3 and TOTO-3 iodide allow detection of microbial cells in soil samples without interference from background fluorescence
Authors: Henneberger R, Birch D, Bergquist P, Walter M, Anitori RP.
Journal: Biotechniques (2011): 190
Preclinical evaluation of novel triphenylphosphonium salts with broad-spectrum activity
Authors: Millard M, Pathania D, Shabaik Y, Taheri L, Deng J, Neamati N.
Journal: PLoS One (2010)