| Value |
15
Min
Range: Table - link Min
|
| Organism |
Bacteria Escherichia coli |
| Reference |
Shaner NC, Steinbach PA, Tsien RY. A guide to choosing fluorescent proteins. Nat Methods. 2005 Dec2(12):905-9. Article link - link Supplementary table 1 PubMed ID16299475
|
| Primary Source |
Shaner NC et al., Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein. Nat Biotechnol. 2004 Dec22(12):1567-72.PubMed ID15558047
|
| Method |
(primary source:) "RFPs were expressed from pBAD
vectors in E. coli LMG194 by growing single colonies in 40 ml RM/ampicillin
supplemented with 0.2% D-glucose for 8 h, adding 40 ml LB/ampicillin and
L-arabinose to a final concentration of 0.2%, and incubating overnight at 37°C.
For maturation experiments, flasks were sealed with parafilm upon induction
to restrict oxygen availability. All proteins were purified by Ni-NTA chromatography
(Qiagen) and dialyzed into PBS. Biochemical and fluorescence characterization
experiments were done as described [ref 5]. For FRET measurements,
purified Zn2+ sensor proteins were diluted in 10 mM MOPS, 100 mM KCl,
pH 7.4 with either 1 mM EDTA or 1 mM ZnCl2, and fluorescence emission
spectra were collected with excitation near the peak donor excitation wavelength." |
| Comments |
For mCherry maturation half-time of ~40min see Khmelinskii et al., 2012 PMID 22729030 p.709 left column 3rd paragraph |
| Entered by |
Uri M |
| ID |
106877 |