AnaSpec is pleased to highlight our wide collection of amine
and thiol reactive 6-isomeric dyes of fluoresceins
and rhodamines, and AnaSpec’s own proprietary HiLyte Fluor™ dyes for labeling
amine and thiol modified oligonucleotides.
Fluorescent dye-labeled oligos are widely used as primers for
sequencing and PCR reactions, and also as probes for fluorescence in situ
hybridization (FISH). Double-labeled oligos are used to produce fluorescence
resonance energy transfer (FRET) for a number of applications such as real-time
PCR assays and automated DNA sequencing.
Synthetic oligos can be modified with amine or thiol groups
at their 5’-end, 3’-end or internally. These amine and thiol incorporated oligos
can then be labeled with amine and thiol reactive fluorophores, respectively.
N-hydroxysuccinimidyl esters (SE) and isothiocyanates of fluorescein
and rhodamine derivatives are used for labeling amine-modified oligos. Maleimides
and iodoacetamides of fluorescein and rhodamine derivatives are used for labeling
thiol-modified oligos.
AnaSpec’s proprietary HiLyte Fluor™ dyes are available in the SE and maleimide
forms. These dyes have fluorescence emission wavelengths spanning the full visible
and near infrared (NIR) spectrum. HiLyte Fluor™ dyes have properties
that are superior to conventional dyes, with high fluorescence quantum yields,
large extinction coefficients, good photostability, low pH sensitivity and improved
water solubility.
Figure 1. The succinimidyl ester (SE) group of the fluorophore
reacts with the amino group of the oligo to form a stable carboxamide bond.
Figure 2. The isothiocyanate group of the fluorophore
reacts with the amino group of the oligo to form a thiourea bond.
Figure 3. The maleimide group of the fluorophore reacts
with the thiol group of the oligo to form a thioether bond.
Figure 4. The iodoacetamide group of the fluorophore reacts
with the thiol group of the oligo to form a thioether bond.
Table 1. Classic dyes for labeling amine- and thiol-modified oligonucleotides
| Product |
Ex/Em |
Size |
Catalog
# |
| 6-FAM, SE
6-Carboxyfluorescein, succinimidyl ester; 6-FAM, NHS ester |
495/517 |
10 mg
100 mg
1 g |
81008
81008-100
81008-1000 |
| 6-FITC
[6-FITC, fluorescein-6-isothiocyanate] |
494/520 |
100 mg |
81010 |
| 6-DTAF
[6-(4,6-Dichlorotriazinyl)aminofluorescein] |
492/517 |
25 mg |
81012 |
| 6-IAF
[6-Iodoacetamidofluorescein] |
483/517 |
25 mg |
81407 |
| 6-TET, SE
6-Carboxy-2; 6-TET, NHS ester |
521/536 |
5 mg |
81022 |
| 6-HEX, SE
6-carboxy-2',4,4',5',7,7'-hexachlorofluorescein, succinimidyl
ester; 6-HEX, NHS ester |
533/550 |
5 mg |
81020 |
| 6-JOE, SE
6-Carboxy-4',5'-dichloro-2',7'-dimethoxy-
fluorescein, succinimidyl ester; 6-JOE, NHS ester |
520/548 |
5 mg |
81011 |
| 6-CR110, SE
6-Carboxyrhodamine 110, succinimidyl ester; 6-CR110, NHS
ester |
498/521 |
5 mg |
81136 |
| 6-CR6G, SE
6-Carboxyrhodamine 6G, succinimidyl ester; 6-CR6G, NHS
ester |
524/551 |
5 mg |
81106 |
| 6-TAMRA, SE
6-Carboxytetramethylrhodamine, succinimidyl ester; 6-TAMRA,
NHS ester |
547/573 |
5 mg
100 mg
1 g |
81126
81126-01
81126-1 |
| Tetramethylrhodamine-6-maleimide
|
542/568 |
5 mg |
81445 |
| Tetramethylrhodamine-6 C2 maleimide
|
544/572 |
5 mg |
81442 |
| 6-TRITC, R isomer
[Tetramethylrhodamine-6-isothiocyanate] |
544/572 |
5 mg |
81152 |
| 6-ROX, SE
6-Carboxy-X-rhodamine, succinimidyl ester; 6-ROX, NHS
ester |
575/602 |
5 mg |
81115 |
| Sulforhodamine 101 sulfonyl chloride
|
588/601 |
10 mg
10g |
81130
inquire |
| Sulforhodamine 101 C2 maleimide
|
588/601 |
5 mg |
81447 |
Table 2. HiLyte Fluor™ dyes for labeling amine- and thiol-modified
oligonucleotides
| Product |
Ex/Em |
Size |
Catalog
# |
| HiLyte Fluor™ 405 acid, SE
HiLyte Fluor™ 405 acid, NHS ester |
404/428 |
1 mg
5 mg |
89317-1
89317-5 |
| HiLyte Fluor™ 488 acid, SE
"HiLyte Fluor™ 488 acid, NHS ester" |
502/527 |
1 mg
5 mg |
81161-1
81161 |
| HiLyte Fluor™ 555 acid, SE
"HiLyte Fluor™ 555 acid, NHS ester" |
552/569 |
1 mg |
81251 |
| HiLyte Fluor? 594 acid, SE
"HiLyte Fluor™ 594 acid, NHS ester" |
593/616 |
1 mg
5 mg |
81272-1
81272-5 |
| HiLyte Fluor™ 647 acid, SE
"HiLyte Fluor™ 647 acid, NHS ester" |
649/674 |
1 mg |
81256 |
| HiLyte Fluor™ 680 acid, SE
"HiLyte Fluor™ 680 acid, NHS ester" |
678/699 |
1 mg |
81261 |
| HiLyte Fluor™ 750 acid, SE
"HiLyte Fluor™ 750 acid, NHS ester" |
754/778 |
1 mg |
81266 |
| HiLyte Fluor™ 405 C2 maleimide
|
404/428 |
1 mg |
89320 |
| HiLyte Fluor™ 488 C2 maleimide
|
502/527 |
1 mg |
81164 |
| HiLyte Fluor™ 555 C2 maleimide
|
552/569 |
1 mg |
81254 |
| HiLyte Fluor™ 594 C2 maleimide
|
593/616 |
1 mg |
81275 |
| HiLyte Fluor™ 647 C2 maleimide
|
649/674 |
1 mg |
81259 |
| HiLyte Fluor™ 680 C2 maleimide
|
678/699 |
1 mg |
81264 |
| HiLyte Fluor™ 750 C2 maleimide
|
754/778 |
1 mg |
81269 |
AnaSpec’s HiLyte
Fluor™ dyes and classic dyes are excellent choices for labeling
amine- and thiol-modified oligonucleotides.
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