
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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1。称量氨基丁三醇 242g,Na2EDTA.2H2O 37.2g 于1L烧杯中;
2。向烧杯中加入约600ml去离子水,充分搅拌均匀;
3。加入57.1ml的冰乙酸,充分溶解;
4。用NaOH调pH至8.3,加去离子水定容至1L后,室温保存。
使用时稀释50倍 即1×TAE Buffer
10×TBE Buffer配制方法:
1。称量氨基丁三醇 108g,Na2EDTA.2H2O 7.44g,硼酸55g 于1L烧杯中;
2。加入约700ml去离子水,搅拌均匀;
3。用NaOH调pH至8.3,加去离子水定容至1L后,室温保存。
使用时稀释10倍 即1×TBE Buffer
蛋白质上样缓冲液中的SDS和DTT分别可以屏蔽蛋白质电荷以及破坏二硫键,避免形成多聚体。
DNA上样缓冲液主要是起沉降和指示作用,不可用于蛋白,否则会影响蛋白相对分子量定量。
同样蛋白上样缓冲液中的tris-HCl浓度过高也会使溴酚蓝条带扭曲。
因为DNA和RNA链上的碱基都基本一致。而这两种上样缓冲液中含有的染料是二甲苯青和溴酚蓝,都可以与其碱基结合,跑出条带。
但是,如果使用DNA上样缓冲液,里面会夹杂一些RNA酶,这些酶有可能会影响到实验结果。使用前需要将RNA酶处理掉。
0.5 M tris-HCl (pH 6.8) 20%
丙三醇 20%
20%SDS 20%
0.1%溴酚兰 5%
2-巯基乙醇 10%
双蒸水 25%
不调PH值,PH在配0.5 M tris-HCl 时已调,缓冲液呈蓝紫色。
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.

