This article from ChemBioChem discusses the use of a modified SNAP tag to study dynamic cellular processes. Background fluorescence was eliminated by simply adding a quencher moiety to the SNAP tag substrate. Other methods to eliminate background fluorescence were time consuming, tedious, or limited downstream research possibilities. The advantage of the SNAP tag system is that media does not have to be removed to see fluorescence; therefore, protein dynamics can be more easily investigated. Live cell imaging and SDS PAGE confirmed the success of using the modified SNAP tag.
Friday, September 23, 2011
Development of SNAP-Tag Fluorogenic Probes for WashFree Fluorescence Imagin
This article from ChemBioChem discusses the use of a modified SNAP tag to study dynamic cellular processes. Background fluorescence was eliminated by simply adding a quencher moiety to the SNAP tag substrate. Other methods to eliminate background fluorescence were time consuming, tedious, or limited downstream research possibilities. The advantage of the SNAP tag system is that media does not have to be removed to see fluorescence; therefore, protein dynamics can be more easily investigated. Live cell imaging and SDS PAGE confirmed the success of using the modified SNAP tag.
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They mentioned that this technology could be applied for HTS experiments. How would this work?
ReplyDeleteI just don't really see how this technique could allow to detect interaction between the protein of interest and a small molecule.
Why can't a terminal cysteine activate this fluorophore, without the bulky snap tag?
ReplyDeleteI guess internal Cys would also work...
Does the Snap tag make it more accessible?
You couldn't use a Cys reactive dye (e.g., an iodoacetamide) intracellularly, since it would label every Cys-containing protein.
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