
Product Name | Thioflavin T *UltraPure Grade* |
Size | 1 g |
Catalog # | AS-88306 |
US$ | $72 |
The benzothiazole dye thioflavin T (ThT) is a classic amyloid stain for senile plaques containing bA4 peptide in Alzheimer"s disease brain. ThT also binds rapidly and specifically to the anti-parallel b-sheet fibrils formed from synthetic b-amyloid (1-40), but does not bind to monomer or oligomeric intermediates. The fibrillar b-sheet-bound dye species undergoes a characteristic 120 nm red shift of its excitation spectrum that may be selectively excited at 450 nm, resulting in a fluorescence signal at 482 nm. ThT is a useful probe for the aggregated fibrillar state of b-amyloid (1-40) fibrils as the amyloid-specific fluorescence reports only fibrillar species. The binding of ThT does not interfere with the aggregation of this peptide into amyloid fibrils. The putative conformational changes detected by the ThT fluorescence suggest that small pharmacologic ligands can perturb and possibly dissociate Ab amyloid fibrils. | |
Detailed Information | ![]() |
Storage | -20°C desiccated and protected from light |
References | 1. Ban T, et al. (2003).J Biol Chem 278, 16462-5. 2. Kung MP, et al. (2002).Brain Res 956, 202-10.3. De Ferrari GV, et al. (2001).J Biol Chem 276, 23282-7. |
Molecular Weight | 318.865 |
Molecular Formula | C17H19ClN2S |
Spectral Properties | Abs/Em = 412/482 nm |
Solvent System | DMSO |
Product Citations | Pronchik, J. et al. (2010). In vitro formation of amyloid from α-synuclein is dominated by reactions at hydrophobic interfaces. J Am Chem Soc 132, 9797.Stsiapura, V. et al. (2010). Charge transfer process determines ultrafast excited state deactivation of thioflavin T in low-viscosity solvents. J Phys Chem A 114, 8345.Sulatskaya, A.I. et al. (2010). Spectral properties and factors determining high quantum yield of thioflavin T incorporated in amyloid fibrils. Spectroscopy 24, 169. |
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我要做的两个目的蛋白,分子量一个是140的,另一个是36,内参用的GAPDH。这几天刚刚要做小鼠组织的蛋白,就把新提取的组织蛋白和之前做过的细胞蛋白一起上了,细胞和组织的各上了3个孔。
电泳转膜孵育等条件都是以前做成熟的
之后显影,细胞蛋白的,两个目的和内参都出得挺好;但是小鼠的组织蛋白只有分子量140的那个显出条带了,36的那个显出来的像一片水渍一样,斑片状的黑,内参有时也这样、有时干脆什么都不显。不知道这是什么原因...按说细胞同样的蛋白做出来,WB的各个步骤应该不会有太大问题的。
有点怀疑蛋白提取的不好,我提组织蛋白的的操作是在冰上进行的,裂解液也加有PMSF,用BCA法测得的蛋白浓度大概也有3~5mg/ml。
在这里求助各位高手~望给我指点一下~不胜感激啊!
去垢剂作为添加剂生长蛋白质晶体是一项很有用的技术。非离子和两性离子结晶剂在膜蛋白结晶中已经得到很好应用,目前已经变成常规的实验手段。去垢剂在一些可溶蛋白中也起到了一定的作用,提高了晶体的质量和结果的可重复性 ,促进了单晶的生长, 但具体的影响机制尚不明确。其可能原因是去垢剂的引入使结晶液滴中的蛋白分子更具有亲水性,使溶液中同质蛋白质分子增加。向左转|向右转
如要膜蛋白保持原来的结构应采用:非离子型去垢剂.
表面活性剂:通常由一个极性头和一个非极性尾组成,用于去除难溶于水的污渍
漂白剂:通常含次氯酸盐成分,原理为其氧化性
稀盐酸:用于去除碱性的污渍,如马桶中的结块
碳酸钠溶液:用于去除油渍,其原理为酯在碱溶液中水解,参见皂化反应

