Tag-lite®

Comprehensive HTRF cellular platform
for cell surface receptor studies and functional assays.

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Tag-lite brings new labeling technology
Tag-lite: a comprehensive selection of tools and reagents streamlined to specifically label fusion protein with synthetic HTRF dyes. Tag-lite combines HTRF with SNAP-tag®, CLIP-tag and HaloTag technologies. This solution is ideal for a wide range of applications, such as mechanistics and receptor dimerization, as well as ligand binding assays and second messenger assessment.

Tag-lite® technology assay principle:

This platform is an original and efficient way to accurately label a protein of interest on a targeted site with HTRF dyes.

Originally developed by the Ecole Polytechnique Federale de Lausanne (EPFL) and commercialized by New England Biolabs, Inc., SNAP-tag® and CLIP-tag are small fusion tags that covalently interact with dyes conjugated to guanine or chloropyrimidine leaving groups via a benzyl linker.

HaloTag, a Promega technology, is another orthogonal system for labeling proteins in living cells and in vitro.

The HaloTag protein is a 33 kDa enzyme derivatized from a prokaryotic hydrolase SNAP-tag®, CLIP-tag and HaloTag can easily be fused to either the N- or the C-terminal position on proteins of interest, and can then be specifically and covalently labeled with their defined substrates.




Tag-lite plasmids

 
To develop your own target

Cisbio Bioassays proposes plasmids encoding SNAP-tag®, CLIP-tag, HaloTag and bordered by restriction sites for cloning a gene of interest.

By using those plasmids, a construction can be engineered with encoding for the protein of interest and SNAP-tag®, CLIP-tag or HaloTag, in the N or C terminal position. Once transfected into cells, this construction leads to the expression of the tagged protein.

Once transfected into cells, this construction leads to the expression of the fused protein of interest with the SNAP-tag®, CLIP-tag or HaloTag. Cisbio proposes a selection of plasmid encoding for a GPCR fused in N-terminal position with SNAP-tag®, CLIP-tag or HaloTag.
Tag-lite® Plasmids MSDS
SNAP-tag® PT8SNAPHYG pT8-SNAP (Hygromycin) on request pdf
PT8SNAPNEO pT8-SNAP (Neomycin) 10 µgr pdf
PT8SNAPZEO pT8-SNAP (Zeocin) on request pdf
CLIP-tag® PT8CLIPHYG pT8-CLIP (Hygromycin) on request pdf
PT8CLIPNEO pT8-CLIP (Neomycin) 10 µgr pdf
PT8CLIPZEO pT8-CLIP (Zeocin) on request pdf
Halotag® PT8HALOHYG pT8-HaloTag (Hygromycin) 10 µgr pdf
PT8HALONEO pT8-HaloTag (Neomycin) 10 µgr pdf
PT8HALOZEO pT8-HaloTag (Zeocin) 10 µgr pdf


SNAP-tag® does not affect the activity of the labelled protein.

 

Reverse transfection of vasopressin 1a receptor was performed with the wild type (WT-V1a) and the SNAP-tag® V1a (ST-V1a) of the receptor. Inositol 1-phosphate production induced by vasopressin stimulation was assessed using the IP-One kit on the two different cellular models.

Studies confirmed that SNAP-tag® fusion on the GPCR N-terminal and its specific labeling with the appropriate fluorescent substrate do not affect either the GPCR binding of a specific ligand or the GPCR function. Vasopressin EC50 measured with IP1 accumulation assay were very similar on both WT-V1a and ST-V1a.


How do the Tag-lite® substrates work?

 

A selection of three substrates is proposed for each SNAP-tag®, CLIP-tag and HaloTag. These substrates are labeled with HTRF fluorophores, terbium cryptate donor (Lumi4®-Tb) and green or red HTRF acceptors.


Tag-lite® substrate chart

 

Description Product reference Quantity MSDS
Tag-lite labeling medium LABMED 100 ml (5X) pdf

Optimized buffer for SNAP- and CLIP-tag labeling.

Fusion HTRF partner Description Product reference Quantity Package insert MSDS
SNAP HTRF donor Tag-lite SNAP-Lumi4Tb SSNPTBC 2 nmoles pdf pdf
SSNPTBD 5 nmoles pdf
SSNPTBG 5x5 nmoles pdf
SSNPTBX 100 nmoles pdf
HTRF acceptors Tag-lite SNAP-Green SSNPGRNE 5 nmoles pdf pdf
SSNPGRNF 5x5 nmoles pdf
SSNPGRNZ 100 nmoles contact us
Tag-lite SNAP-Red SSNPREDE 20 nmoles pdf pdf
SSNPREDF 5x20 nmoles pdf
SSNPREDZ 500 nmoles contact us
CLIP HTRF donor Tag-lite CLIP-Lumi4Tb SCLPTBE 20 nmoles pdf pdf
SCLPTBF 5x20 nmoles pdf
SCLPTBZ 500 nmoles contact us
HTRF acceptors Tag-lite CLIP-Green SCLPGRNE 20 nmoles pdf pdf
SCLPGRNF 5x20 nmoles pdf
SCLPGRNZ 500 nmoles contact us
Tag-lite CLIP-Red SCLPREDE 20 nmoles pdf pdf
SCLPREDF 5x20 nmoles pdf
SCLPREDZ 500 nmoles contact us
HaloTag HTRF donor HaloTag-Lumi4®-Tb SHALOTBC 2 nmoles pdf pdf
HTRF acceptors HaloTag-Green SHALOGRNE 5 nmoles pdf pdf
HaloTag-Red SHALOREDE 5 nmoles pdf pdf