What does the SNAP-tag do?

SNAP-tag is a self-labeling protein derived from human O6-alkylguanine-DNA-alkyltransferase. SNAP -Tag reacts with covalently with O6-benzylguanine derivatives, for example fluorescent dyes conjugated to guanine or chloropyrimidine. It can be used as a protein tag for tagging your protein of interest (POI).

What is snap biochemistry?

SNAP-tag® is a self-labeling protein tag commercially available in various expression vectors. SNAP-tag is a 182 residues polypeptide (19.4 kDa) that can be fused to any protein of interest and further specifically and covalently tagged with a suitable ligand, such as a fluorescent dye.

What do snare proteins do?

The primary role of SNARE proteins is to mediate vesicle fusion – the fusion of vesicles with the target membrane; this notably mediates exocytosis, but can also mediate the fusion of vesicles with membrane-bound compartments (such as a lysosome).

What is GFP tagging?

GFP-tagging is a way of preparing a sample for fluorescence microscopy by using the GFP as a fluorescent protein reporter. This is done by cloning the GFP in frame with the target protein at either the N- or C-terminus of the amino acid chain.

Why is GFP used?

Biologists use GFP as a marker protein. GFP can attach to and mark another protein with fluorescence, enabling scientists to see the presence of the particular protein in an organic structure. Gfp refers to the gene that produces green fluorescent protein.

What are the T SNARE proteins?

The SNARE proteins, syntaxin, SNAP-25, and synaptobrevin have long been known to provide the driving force for vesicle fusion in the process of regulated exocytosis.

Where do you tag proteins?

They can be added to either end of the target protein, so they are either C-terminus or N-terminus specific or are both C-terminus and N-terminus specific. Some tags are also inserted into the coding sequence of the protein of interest; they are known as internal tags.

What is the CLIP-tag and how do I use it?

If you’re looking to do dual labeling, then the CLIP-tag is for you. The CLIP-tag was developed to complement the SNAP-tag so that you can apply both types of fusions in one experiment. The CLIP-tag protein is also a derivative of hATG, so it shares the features of its sibling, the SNAP-tag protein.

What are snap-and CLIP-tag protein labeling?

Protein Labeling (SNAP/CLIP) SNAP- and CLIP-tag protein labeling systems enable the specific, covalent attachment of virtually any molecule to a protein of interest. There are two steps to using this system: cloning and expression of the protein of interest as a SNAP-tag ® fusion, and labeling of the fusion with the SNAP-tag substrate of choice.

What is a CLIP peptide?

CLIP or Class II-associated invariant chain peptide is the part of the invariant chain (Ii) that binds to the peptide binding groove of MHC class II and remains there until the MHC receptor is fully assembled. CLIP is one of the most prevalent self peptides found in the thymic cortex of most antigen-presenting cells.

What is the difference between CLIP-tag™ and SNAP-tag?

In the labeling reaction, the substituted benzyl group of the substrate is covalently attached to the SNAP-tag. CLIP-tag™ is a modified version of SNAP-tag, engineered to react with benzylcytosine rather than benzylguanine derivatives.