| Oligomerization | Organism | Molecular Weight | Cofactor |
|---|---|---|---|
| Monomer | Discosoma sp. | 27.0 kDa | - |
| Ex λ | Em λ | EC (M-1 cm-1) | QY | Brightness | pKa | Maturation (min) | Lifetime (ns) |
|---|---|---|---|---|---|---|---|
| 568 | 592 | 75,000 | 0.49 | 36.75 | 6.5 | 30.0 | 2.9 |
| % Normal Cells | OSER/NE ratio | Cell Type | Reference |
|---|---|---|---|
| 95.3 ± 1.7 (10000 cells) | - | HeLa | Cranfill et al. (2016) |
| 87.0 (103 cells) | 2.2 ± 0.2 (14 cells) | U-2 OS | Bindels et al. (2016) |
| t1/2 (s) | Power | Light | Mode | In Cell | Fusion | ˚C | Reference |
|---|---|---|---|---|---|---|---|
| 4.8 | Bindels et al. (2016) |
mApple was derived from mOrange with the following mutations: R17H/G40A/T66M/A71V/V73I/K92R/V104I/V105I/T106H/T108N/E117V/S147E/G159S/M163K/T174A/S175A/G196D/T202V
amino acid numbers relative to DsRed. show relative to mOrange
Six generations of directed evolution [of mOrange] with constant photostability selection yielded the variant mApple, which, though substantially brighter than mCherry, displayed complex photoswitching behavior. This behavior was more pronounced with continuous wide-field than with laser-scanning illumination and could be largely eliminated by excitation at alternate wavelengths or by intermittent illumination. However, given our later results using the brighter TagRFP as starting material, we chose not to pursue mApple any further.
Shaner et al. (2008)
Negligible photochromic behavior was measured for the mScarlet variants, while TagRFP-T, mRuby2, mRuby3 and mApple showed 15%, 19%, 41%, and 51% photochromic behavior, respectively. Hence, extreme care must be taken when using the latter four RFP variants as acceptors in FRET studies, since a photochromic effect is easily confused with a changed FRET state, especially if one considers that the typical FRET contrast in many sensors is in the range of only 5–20%. The photochromic behavior can also interfere with characterization of FPs, like determination of photostability (Supplementary Fig. 8) or brightness (Supplementary Fig. 5l).
Bindels et al. (2016)
(2008). Nature Methods, 5(6) , 545-551. doi: 10.1038/nmeth.1209. Article Pubmed
(2016). Nature Methods, 14(1) , 53-56. doi: 10.1038/nmeth.4074. Article Pubmed
Something missing or incorrect? Submit a change Submit a change