
Product Name | HiLyteᵀᴹFluor 647 acid, SEHiLyte Fluor 647 acid, NHS ester |
Size | 1 mg |
Catalog # | AS-81256 |
US$ | $249 |
Purity | ≥95% by HPLC |
HiLyte Fluor 647 acid, SE is an excellent fluorescent labeling dye that generates the protein conjugates that are slightly red-shifted compared to those of Cy5 dyes, resulting in an optimal match to filters designed for Cy5 dyes. However, total fluorescence of the secondary antibody conjugates of HiLyte Fluor 647 dyes is significantly higher than that of Cy5 conjugates supplied by other companies. Also, unlike Cy5 dyes, HiLyte Fluor 647 dyes have very little change in absorption or fluorescence spectra when conjugated to most proteins, oligonucleotides and nucleic acids, thus yielding greater total fluorescence at the same degree of substitution. Additionally, our in-house data indicated that HiLyte Fluor 647 is superior to Cy5 in fluorescence polarization-based assays.Extinction Coefficient (M-1cm-1): 250,000Fluorescence quantum yield: 0.33Fluorescence Life Time (ns): 1.0 | |
Detailed Information | ![]() |
Storage | -20°C desiccated and protected from light |
Molecular Weight | 1302.71 |
Spectral Properties | Abs/Em = 649/674 nm |
Solvent System | DMF or DMSO |
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最好是测浓度,一般不超过30-40ug,上样缓冲液一般4×或者5×,这个没太大区别。
蛋白电泳上样缓冲液包括2%SDS , 0.1%溴酚蓝 10%甘油,SDS是保证样品中的所有蛋白带电荷一致,减少电荷对电泳结果的影响。还原剂是让二硫键处于断开状态,保证蛋白分子的线性.
溴酚蓝作用是指示上样蛋白的跑胶时标记的,甘油是增加上样重量的,以免漂浮,煮沸是使蛋白变性的永久保存
加样缓冲液的主要作用是使PCR产物与其混合,
使DNA沉于加样孔的底部,防止DNA跑出来.
2乘SDS-PAGE上样缓冲液(20ml体系)
1.0mol/L Tris-HCL (pH 6.8) 2ml
1.0mol/L DTT 4ml
SDS 0.8g
溴芬蓝 0.04g
甘油 4ml
dd水 定容至20ml
4℃冰箱保存,可以不分装,我们实验室一般用三个月以上
先配1.0mol/L的DTT,双蒸无菌水溶解4℃冰箱保存,配上样缓冲液的时候加入就可以了,都混匀保存也没问题的

