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- 文献和实验
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- 库存:
大量
- 供应商:
北京百奥创新科技有限公司
- 规格:
5mL;50mL;1L
胺基活化琼脂糖凝胶6HF
产品名称:胺基活化琼脂糖凝胶6HF英文名称:Amine-activated SepFast
Product Introduction:
Amine-activated agarose beads can be used for the immobilization of carboxyl containing or aldehyde containing molecules. The coupling chemistry is carbodiimide-based and is a successful and well-documented technique. Amine-activated SepFast forms a chemically stable amide linkage with ligands containing carboxyl groups. It has a long spacer (9 atoms). The pre-activated agarose base matrix can be readily employed to make various custom affinity chromatography adsorbents for both small scale and large scale purification applications.

Product Properties:
Amine-activated SepFast 4 High Flow (4HF) is made of highly cross-linked 4% beaded agarose. It shows high mechanical rigidity allowing high flow throughput with reduced back pressure.
Agarose has long been used for chromatographic separations due to its excellent hydrophilic and low non-specific-binding nature. The particles have an open pore structure with excellent mass transfer properties to large protein molecules.
The base matrix is activated by attaching a long hydrophilic chain with an amine group in the end. They react with either carboxyl containing ligands in the presence of a carbodiimide agent, or aldehyde containing ligands in the presence of a reducing agent. Amine-activated SepFast is supplied as an aqueous suspension in 20% ethanol.
Product Series:
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文献和实验S.L. Lim, H.W. Ng, M.A. Akwiditya, C.W. Ooi, E.S. Chan, K.L. Ho, W.S. Tan, G.K. Chua, B.T. Tey, Single-step purification of recombinant hepatitis B core antigen Y132A dimer from clarified Escherichia coli feedstock using a packed bed anion exchange chromatography, Process Biochemistry, 2018, 69:208-215.
化学试剂活性愈高,失活率愈大。为克服这个问题,制造商现在采用对目标探针特异性反应的低活性化学基团。醛基和胺基间的反应就是个典型例子,其中胺基和醛基的功能性都比较稳定。另一个问题是非特异性结合。活化膜在应用前必须进行封闭,否则会有额外的样品附着在膜上。封闭步骤需简单、快速、高效。理想的封闭结果是形成一个疏水、无电荷的膜表面,然而通常情况下这很难做到。靶物质和膜结合后也必须进行封闭。膜表面钝化将引入一个新的化学反应,虽然这些新的化学反应不和核酸分子作用,但它可增加膜表面的电荷及疏水性进而导致非特异
,那就要考虑一下这个蛋白是不是二聚体或者多聚体啦。▪蛋白修饰要变大如果排除了以上 4 条原因,预测值和实验结果不一致,那就是因为我们面对的蛋白质,是个善变的对象,它不仅会剪切活化,还有在翻译表达后修饰。比如:比如甲基化、乙酰化、磷酸化、糖基化、烷基化、生物素化、谷氨酸化、甘氨酸化、硫酸化、异.戊二烯化、硫辛酸化、磷酸泛酰巯基乙.胺基化……balabala。在不同状态下的蛋白质还会有不同的修饰。加入了官能团肯定会让蛋白质比预测值偏大。在日常的实验中,糖基化是比较常见的修饰,一般会导致跑出来的条带比预测
性好、孔径均匀适当、宜于活化等优点,因此得到了广泛的应用。琼脂糖凝胶微球的商品名为Sepharose,含糖浓度为2%、4%、6%时分别称为2B、4B、6B。因为Sepharose 4B的结构比6B疏松,而吸附容量比2B大,所以4B应用最广。基质的活化基质的活化是指通过对基质进行一定的化学处理,使基质表面上的一些化学基团转变为易于和特定配体结合的活性基团。配体和基质的偶联,通常首先要进行基质的活化。多糖基的活化多糖基质尤其是琼脂糖是一种常用的基质。琼脂糖通常含有大量的羟基,通过一定的处理可以引入各种适宜






