供体的发射光谱与受体的吸收光谱有重叠;
供体和受体之间的距离在1 - 10纳米(1纳米=10埃,C-C单键长为1.5埃左右)范围内;
供体和受体的跃迁偶极具有适当的相对取向。
酶活性检测;
酶药物靶点的筛选和发现;
酶抑制剂高通量筛选。
供体(荧光染料)
Trp (Tryptophan)
Trp (Tryptophan)
Mca (7-Methoxycoumarin-4-yl)acetyl)
Abz (2-Aminobenzoyl)
Abz (2-Aminobenzoyl)
Abz (2-Aminobenzoyl)
Abz (2-Aminobenzoyl)
Abz (2-Aminobenzoyl)
N-Me-Abz(N-Methyl-anthraniloyl)
EDANS (5-[(2-Aminoethyl) amino] naphthalene-1-sulfonic acid)
5-FAM (5-Carboxyfluorescein)
5-FAM (5-Carboxyfluorescein)
CP488
Lucifer Yellow
FITC (Fluorescein isothiocyanate)
Dansyl (5-(Dimethylamino) naphthalene-1-sulfonyl)
5-TAMRA (Carboxytetramethyl rhodamine)
5-TAMRA (Carboxytetramethyl rhodamine)
Eu(III) Chelate
Cy5
Cy5.5
颜色
最大吸收波长
280nm
280nm
325nm
320nm
320nm
320nm
320nm
340nm
340nm
430nm
492nm
492nm
495nm
430nm
490nm
342nm
547nm
547nm
340nm
647nm
648nm
最大发射波长
360nm
360nm
392nm
420nm
420nm
420nm
420nm
420nm
440nm
490nm
512nm
512nm
519nm
520nm
520nm
562nm
573nm
573nm
613nm
665nm
701nm
消光系数M-1 cm-1 (ε)
5,600
5,600
11820
/
/
/
/
/
/
5,900
83,000
/
/
11,000
73,000
3,300
90,000
/
/
/
/
受体(淬灭基团)
Dnp (2,4-Dinitrophenyl)
4-Nitro-Z (4-Nitro-benzyloxycarbonyl)
Dnp (2,4-Dinitrophenyl)
Dnp (2,4-Dinitrophenyl)
EDDnp(N-(2,4-Dinitrophenyl)ethylenediamine)
pNA (para-Nitroaniline)
3-Nitro-Tyr (3-Nitro-tyrosine)
4-Nitro-Phe (4-Nitro-phenylalanine)
Dnp (2,4-Dinitrophenyl)
Dabcyl (4-(4-Dimethylaminophenylazo)benzoyl)
CPQ2
Dabcyl (4-(4-Dimethylaminophenylazo)benzoyl)
CPQ2
Dabsyl (4-(4-Diethylaminophenylazo)-benzenesulfonyl)
Dnp (2,4-Dinitrophenyl)
4-Nitro-Phe (4-Nitro-phenylalanine)
QSY7
BHQ3
QSY-7
QSY21
QSY21