CLIP-Vista Green is a green fluorescent substrate that can be used to label CLIP-tag™ fusion proteins (in cell lysates or purified proteins) for detection by SDS-PAGE. This substrate (BC-Vista Green) is based on fluorescein and is optimized for excitation with the 488 nm laser excitation line in a laser based gel scanner. It can also be excited using 360 nm light from a standard UV-transilluminator. It has an excitation maximum at 500 nm and emission maxima at 524 nm. This package includes 50 nmol of CLIP-Vista Green substrate, sufficient to label one hundred 20 µl samples containing CLIP-tag fusion protein for in-gel detection.
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 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, and labeling of the fusion with the CLIP-tag substrate of choice. Expression of CLIP-tag fusion proteins is described in the documentation supplied with CLIP-tag plasmids. The labeling of the fusion proteins with the CLIP-tag substrate for detection by SDS-PAGE is described in this document.
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