Custom assay development

cat.#AD001

Fluorescent proteins (FPs) act as endogenously expressed biolabels for fluorescence microscopy that makes them particularly suitable for use in various HTS/HCS-type, live-cell, drug-discovery screening assays. Red and far-red FPs, which are easily distinguished from cellular autofluorescence and from the typical fluorescence wavelength spectrum of many medicinal chemistry compounds, provide technologically useful possibilities to design of sophisticated, novel assay formats for drug discovery.
Selection and evaluation of an appropriate FP marker along with optimization of assay parameters is a laborious and time consuming procedure.

We offer the following custom research services to assist with FP selection / assay development:

    1. Making an agreed number of different constructs of interest (FP--linker--target protein)
    * advice on the fuse and/or linker structure is an option
    2. Testing the constructs in an number of model cell lines to find the best ones.
    3. Testing the selected constructs in various cell lines to select the most appropriate cells in terms of fusion localization/redistribution.
    4. Making stably transfected cell lines based on the testing results above.

For customers, who wish to multiplex HCS readout to achieve real multiparameter imaging, we offer unique option to fuse protein of interest with bright monomeric red (TagRFP) and far-red (TagFP635) fluorescent proteins that could be used in combination with dyes/FPs visible in UV, blue and green part of spectrum. Both proteins perfectly fit for fusions.

TagRFP use for cell, cellular organelle, and protein labeling.

Microscopic images of HeLa cells transiently transfected with TagRFP and TagRFP-tagged fusions. Images were kindly provided by Michael W. Davidson (Florida State University).

TagRFP description

TagFP635 use for protein labeling.

Microscopic images of HeLa cells transiently transfected with TagFP635-tagged fusions.
Images were kindly provided by Michael W. Davidson (Florida State University).

TagFP635 description

Related publications:

  • Belousov VV, Fradkov AF, Lukyanov KA, Staroverov DB, Shakhbazov KS, Terskikh AV, Lukyanov S. Genetically encoded fluorescent indicator for intracellular hydrogen peroxide. Nat Methods. 2006; 3 (4):281-6. / pmid: 16554833
  • Chudakov DM, Lukyanov S, Lukyanov KA. Fluorescent proteins as a toolkit for in vivo imaging. Trends Biotechnol. 2005 Dec;23(12):605-13. / pmid: 16269193
  • Merzlyak EM, Goedhart J, Shcherbo D, Bulina ME, Shcheglov AS, Fradkov AF, Gaintzeva A, Lukyanov KA, Lukyanov S, Gadella TW, Chudakov DM. Bright monomeric red fluorescent protein with an extended fluorescence lifetime. Nat Methods. 2007; 4 (7):555-7. / pmid: 17572680
  • Shcherbo D, Merzlyak EM, Chepurnykh TV, Fradkov AF, Ermakova GV, Solovieva EA, Lukyanov KA, Bogdanova EA, Zaraisky AG, Lukyanov S, Chudakov DM. Bright far-red fluorescent protein for whole-body imaging. Nat Methods. 2007; 4 (9):741-6. / pmid: 17721542
  • Souslova EA, Belousov VV, Lock J, Stromblad S, Kasparov S, Bolshakov AP, Pinelis VG, Labas YA, Lukyanov S, Mayr LM, Chudakov DM. Single fluorescent protein-based Ca2+ sensors with increased dynamic range. BMC Biotechnol. 2007; 7 :37. / pmid: 17603870
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