Dyes for labeling Amine- and Thiol-Modified Oligos


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 premium quality and cost effective HiLyte™ Fluor dyes and classic dyes are excellent choices for labeling amine- and thiol-modified oligonucleotides.