Biology Reference
In-Depth Information
Table 1 Overview of the spectral properties of the noncanonical FP variants. Additional values in
brackets were taken from [
12
]
Variant
Absorbance
maximum/
nm
Extinction
coefficient/
M
1
cm
1
Emission
maximum/
nm
Quantum
yield
Relative
fluorescence
Ref.
CFP6
449
25,900
1,000
488
0.140
-
[
13
]
CFP6[(6-Cl)Trp]
430
19,800
6,400
474
0.050
-
[
13
]
CFP6[(6-Br)Trp]
430
20,600
3,800
474
0.058
-
[
13
]
CFP6[BT]
415
8,100
1,100
472
0.048
-
[
13
]
GFPuv
397/475
25,000
2,000/
6,000
506
0.760
-
[
14
]
500
GFPuv[(4-Am)Phe] 435
31,000
4,000
498
0.43
-
[
14
]
GFPuv[(4-Me)Tyr]
394
27,000
1,000
460
0.37
-
[
14
]
GFPuv[(4-Br)Phe]
375
20,000
1,000
428
0.014
-
[
14
]
GFPuv[(4-I)Phe]
381
16,000
1,000
438
0.013
-
[
14
]
ECFP
434
24,800 (20,000)
476
(0.15)
1
[
15
]
ECFP[(4-F)Trp]
426
14,810
697
470
-
0.4
[
15
]
ECFP[(5-F)Trp]
426
19,172 730
466
-
0.7
[
15
]
ECFP[(6-F)Trp]
432
21,762
238
474
-
0.5
[
15
]
16,650
837
ECFP[(7-F)Trp]
426
471
-
0.4
[
15
]
ECFP[(4-Me)Trp]
438
23,450
561
480
-
1
[
15
]
ECFP[(4-Am)Trp]
466
23,700
574
0.15
-
[
11
],
[
70
]
ECFP[[2,3]Sep]
435
4,533
239
-
-
-
[
15
]
ECFP[[3,2]Sep]
388
11,650
975
-
-
-
[
15
]
ECFP[[2,3]Tpa]
385
6,840
421
-
-
-
[
15
]
ECFP[[3,2]Tpa]
388
6,350
717
-
-
-
[
15
]
EGFP
488
35,570
1,316
510
(0.7)
1.00
[
15
]
(23,000)
EGFP[[2,3]Sep]
488
40,520
1,261
509
-
0.9
[
15
]
EGFP[[3,2]Sep]
488
49,930
1,597
509
-
0.83
[
15
]
EGFP[[2,3]Tpa]
488
30,715
1,076
509
-
1.25
[
15
]
43,715
1,455
EGFP[[3,2]Tpa]
488
509
-
0.83
[
15
]
ds
Red
556
59,000 (27,300)
603 (592)
0.04
(0.14)
-
[
16
]
ds
Red[(3-Am)Tyr]
556
59,000
615
0.16
-
[
16
]
ds
Red[(3-F)Tyr]
556
47,000
591
0.25
-
[
16
]
EGFP[(2-F)Tyr]
482
22,190
1,174
504
-
0.6
[
17
]
EGFP[(3-F)Tyr]
485
30,200
1,200
514
-
0.9
[
17
]
EYFP
514
48,770
1,250
(45,000)
527
(0.54)
1
[
17
]
EYFP[(2-F)Tyr]
504
55,460
2,180
520
-
1.2
[
17
]
EYFP[(3-F)Tyr]
518
48,790
1,440
533
-
1
[
17
]
of
100 nm. A comparable Stokes shift could only be detected in class I and III
av
GFPs when the chromophore is excited in its neutral form at 395 nm and
fluoresces from its anionic form at 511 nm. However, the high Stokes shift of
av
GFP occurs due to ESPT [
3
,
19
]. In GdFP, time-resolved experiments do not give
any indication for ESPT [
11
].
A possible explanation for the large Stokes shift could be that, in contrast to
ECFP, the dipole moment of the GdFP chromophore is considerably larger in the
excited state than in the ground state. The additional amino group attached to the
aromatic chromophore moiety has a great influence on the nature of the excited
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