Thioflavine S (Thioflavin S, Direct Yellow 7) is a fluorogenic compound which becomes fluorescent only after oxidation. Thioflavine S is used as a fluorescent histochemical marker of dense core senile plaques.Thioflavine S can bind to dense-core amyloid-β deposits.
| 规格 | 价格 | 库存 | 数量 | 操作 |
|---|---|---|---|---|
| 100mg | ¥446.00 | 现货 | 1 |
Thioflavine S (Thioflavin S, Direct Yellow 7) is a fluorogenic compound which becomes fluorescent only after oxidation. Thioflavine S is used as a fluorescent histochemical marker of dense core senile plaques.Thioflavine S can bind to dense-core amyloid-β deposits.
[1] McLellan ME, et al. J Neurosci. 2003 Mar 15;23(6):2212-7. [1] Bacskai B J, et al. Nature medicine, 2001, 7(3): 369-372.
本方案仅提供一个指导，请根据您的具体需要进行修改。
1.制备染色液：
(1)细胞实验染液：将Thioflavine S溶解于1xPBS中，得到浓度为0.1mg/ml的染料工作液；
(2)组织切片染液：将Thioflavine S溶解于50%乙醇中，得到浓度为0.125mg/ml的染料工作液。
注意: 请根据实际情况调整工作液浓度，现用现配。
2.细胞染色
(1)在无菌盖玻片上培养贴壁细胞；
(2)从培养基中移走盖玻片，使用1x PBS清洗1-2次；
(3)使用 4％多聚甲醛固定细胞，室温固定20min；
(4)从盖玻片的一角加入100μL的染料工作液，轻轻晃动使染料均匀覆盖所有细胞。37 °C避光孵育5-30分钟。不同细胞最佳孵育时间不同，请根据具体实验需求自行摸索。
(5)孵育结束后吸弃染料工作液，使用1x PBS清洗盖玻片1-2次。
3.组织切片染色
(1) Thioflavine S适用于冷冻切片染色和石蜡切片染色；
(2) 将处理后的切片在Thioflavine S的组织切片染液中染色3-10分钟。请根据具体实验需求自行摸索染色时间；
(3) 将切片在50%乙醇和H2O中分别漂洗两次；
4.显微镜检测：Thioflavin S的最大激发光和发射光波长分别为391/428nm。
注意事项：
1）荧光染料均存在淬灭问题，请尽量注意避光，以减缓荧光淬灭。
2）为了您的安全和健康，请穿实验服并戴一次性手套操作。
References:
[1]. Megan E McLellan,et al. In vivo imaging of reactive oxygen species specifically associated with thioflavine S-positive amyloid plaques by multiphoton microscopy. 2003 Mar 15;23(6):2212-7. doi: 10.1523/JNEUROSCI.23-06-02212.2003.
[2].Anyang Sun, Xuan V Nguyen, Guoying Bing.Comparative analysis of an improved thioflavin-s stain, Gallyas silver stain, and immunohistochemistry for neurofibrillary tangle demonstration on the same sections. 2002 Apr;50(4):463-72. doi: 10.1177/002215540205000403.
[3].R Guntern, C Bouras, P R Hof, P G Vallet. An improved thioflavine S method for staining neurofibrillary tangles and senile plaques in Alzheimer's disease. 1992 Jan 15;48(1):8-10. doi: 10.1007/BF01923594.
