Protein localization in live cells


Determine the subcellular localization of a protein by fluorescent imaging.


How it works:
The protein of interest is expressed as a SNAP-tag fusion in the appropriate host cell. The protein is labeled done in living cells using cell-permeable SNAP-cell substrates, or in fixed cells using selected SNAP-vitro substrates. Localization is determined by fluorescent microscopy.
Advantages:
- Easy change of fluorophore.
- Remains stable after fixation.
- Labeling after fixation possible.

 

 

Cytochrome oxidase 8-2-SNAP labeled with BG-DAF (green) in mitochondria of CHO-K1 cells, counterstained with Draq5 (red).

Calnexin-SNAP labeled with TMR-Star (red) in endoplasmic reticulum of U-2 OS cells, counterstained with Hoechst 33342 (blue).

 
Products:

Cell permeable
SNAP-cell 430
SNAP-cell DAF
SNAP-cell 505
SNAP-cell TMR-Star

Cell impermeable
SNAP-vitro 488
SNAP-vitro 547
SNAP-vitro 647



Literature:

Application notes:
SNAP-tag fusion proteins localize correctly in cultured mammalian cells
SNAP-tag fluorescent labeling is stable to various fixation conditions
SNAP-vitro for labeling SNAP-tag fusion proteins in fixed CHO-K1 cells
Correct Localization of SNAP-tag Fusion Proteins in Subcellular Organelles

Publications:
Pick et al.: "Distribution Plasticity of the Human Estrogen Receptor alpha in Live Cells: Distinct Imaging of Consecutively Expressed Receptors". J Mol Biol. (October 10, 2007), /Epub ahead of Print)
Keppler A. et al.: "Fluorophores for live cell imaging of AGT fusion proteins across the visible spectrum" BioTechniques (August 2006), Vol 41, Nr 2 (E pub ahead of print).
Gronemeyer T. et al.:"Directed evolution of O6-alkylguanine-DNA alkyltransferase for applications in protein labeling" Prot Eng Des Sel (2006) (E pub ahead of print).
Regoes A. et al.:"SNAP-tag mediated live cell labeling as an alternative to GFP in anaerobic organisms." Bio Techniques (December 2005), Volume 39, Number 6, pp. 809-812
Keppler A. et al.:"Labeling of fusion proteins with synthetic fluorophores in live cells." PNAS 101 (2004), pp. 9955-9959.
Keppler A. et al.:"Labeling of fusion proteins of O(6)-alkylguanine-DNA alkyltransferase with small molecules in vitro and in vivo." Methods 32 (2004), pp. 437-444.
Keppler A. et al.:"A general method for the covalent labeling of fusion proteins with small molecules in vivo." Nature Biotechnology 21 (2003), pp.86-89.
Juillerat A. et al.:"Directed evolution of O(6)-alkylguanine-DNA alkyltransferase for efficient labeling of fusion proteins with small molecules in vivo." Chem. Biol. 10 (2003), pp.313-317.