NEB/CLIPCell™ 505/S9217S/50 nmol

Description:

CLIP-Cell™505is a green fluorescent substrate that can be used to label CLIP-tag™fusion proteins inside living cells, on cell surfaces or in vitro. This cell-permeable substrate (BC-505) is based on the Dyomics dye DY-505 and is suitable for standard fluorescein filter sets. It has an excitation maximum at 504 nm and emission maximum at 532 nm. 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-DNAalkyltransferase (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-NK1R

Figure 2: Excitation (dotted line) and emission spectra of label 505 coupled to CLIP-tag in buffer at pH 7.5

Notes:

Storage: CLIP-Cell 505 should be stored at -20°C (long term) or at 4°C in the dark (short term, less than 4 weeks). Protect the substrate from light and moisture. With proper storage at -20°C the substrate should be stable for at least three years dry or 3 months dissolved in DMSO.