This package includes 50 nmol of CLIP-Cell 505 substrate, sufficient to make 10 ml of a 5 µM CLIP-tag fusion protein labeling solution.
The CLIP-tag protein labeling system enables the specific, covalent attachment of virtually any molecule to a protein of interest. CLIP-tag is a protein tag based on human O6-alkylguanine-DNA-alkyltransferase (hAGT). CLIP-tag substrates are derivatives of benzylcytosine (BC). In the labeling reaction, the substituted benzyl group of the substrate is covalently attached to the reactive cysteine of CLIP-tag forming a stable thioether bond. Although CLIP-tag is based on the same protein as SNAP-tag, the benzylcytosine substrates form a separate class of substrates, different from the benzylguanine substrates recognized by SNAP-tag. CLIP-tag and SNAP-tag can be used for orthogonal and complementary labeling of two proteins simultaneously in the same cells.
There are two steps to using this system: subcloning and expression of the protein of interest as a CLIP-tag fusion with the CLIP-tag fusion proteins is described in the documentation supplied with CLIP-tag plasmids. The labeling of the fusion proteins is described in this document. Figure 1:Live CHO-K1 cells transiently transfected with pCLIP-NK1RCells were labeled with CLIP-Cell 505 (green) for 60 minutes and counterstained with Hoechst 33342 (blue).Figure 2:Excitation (dotted line) and emission spectra of label 505 coupled to CLIP-tag in buffer at pH 7.5Figure 3:Structure of CLIP-Cell 505 (MW 586.6 g/mol)
This product is related to the following categories:
CLIP-tag™ Substrates Products,
Protein Labeling Products
This product can be used in the following applications:
同源重组是基因组操作中最精确的一种方法。作为一个工具,同源重组已经广泛地用于细菌、酵母、鼠胚胎干细胞(E S ) , 以及其他一些特殊细胞系,但在其他系统如哺乳动物体细胞遗传学上还没有得到应用。最近的研究工作表明,在哺乳动物体细胞中,双链 D N A 发生基因特异的断裂,能够引起上千倍的同源重组。目前,利用锌指核酸酶(Z F N ),能够在哺乳动物基因组中形成这种双链断裂。 Z F N 是一种人工蛋白质,由一个铸指D N A 结合结构域与一个非特异性的核酸酶结构域融合而成。本章描述如何鉴定Z F N 切割 查看更多>