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 technology. 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:

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. SNAP-tag and CLIP-tag 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. In combination with Tag-lite substrates, this platform is an original and efficient way to accurately label a protein of interest on a targeted site with HTRF dyes.




Tag-lite plasmids

 
To develop your own target

Cisbio Bioassays proposes plasmids encoding SNAP- or CLIP-tag 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- or CLIP-tag in the N or C terminal position (1). Once transfected into cells, this construction leads to the expression of the fused protein of interest with the SNAP- or CLIP-tag (2).

Once transfected into cells, this construction leads to the expression of the fused protein of interest with the SNAP- or CLIP-tag (2). Cisbio proposes a selection of plasmid encoding for a GPCR fused in N-terminal position with SNAP- or CLIP-tag.
Tag-lite® Plasmids Sequence
Tag-lite SNAP Plasmids PSNPPOS Tag-lite pSNAP(+) 10 µgr pdf TXT format Gene bank format
PSNPNEG Tag-lite pSNAP(-) 10 µgr pdf TXT format Gene bank format
Tag-lite CLIP Plasmids PCLPPOS Tag-lite pCLIP(+) 10 µgr pdf TXT format Gene bank format
PCLPNEG Tag-lite pCLIP(-) 10 µgr pdf TXT format Gene bank format


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- or CLIP-tag. 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
Tag-lite labeling medium LABMED 100 ml (5X)

Optimized buffer for SNAP- and CLIP-tags labeling.

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