Tide Quenchers, the Best Dark FRET Acceptors Covering the Full Visible and NIR Spectrum
Tide Quencher™ Acceptor Dyes, Optimized to Maximize FRET Efficiency
Although DABCYL has been used to develop a variety of FRET applications, its low quenching efficiency of longer wavelength dyes (such as fluoresceins, rhodamines and cyanines) has limited its use in the development of sensitive fluorogenic FRET probes. Additionally, the absorption spectrum of DABCYL is environment-sensitive. AAT Bioquest has developed the robust Tide Quencher™ acceptor dyes for the development of longer wavelength FRET probes. These Tide Quencher™ dark FRET acceptors (such as TQ1, TQ2, TQ3, TQ4, TQ5, TQ6 and TQ7) are optimized to pair with our Tide Fluor™ dyes and the classic fluorophores (such as AMCA, EDANS, FAM, TAMRA, HEX, JOE, TET, ROX, Cy3, Cy5 and Cy7). Like our Tide Fluor™ donor dyes, our Tide Quencher™ acceptor dyes are much more cost-effective with comparable or even better performance for your desired biological applications than other similar products on the market.
The internally quenched FRET peptide substrate is digested by a protease to generate the highly fluorescent peptide fragment. The fluorescence increase is proportional to the protease activity.
HIV Protease cleavage of Arg-Glu(5-FAM)-Val-Ser-Phe-Asn-Phe-Pro-Gln-Ile-Thr-Lys(TQ2)-Arg. The substrate is incubated with HIV protease . Upon HIV protease cleavage, the fluorescence of 5-FAM is recovered, and monitored with Ex/Em = 490 nm/520 nm.
The Advantages of Tide Quencher™ Dyes:
- Most Powerful: TQ dyes enable you to explore the FRET potentials that might be impossible with other quenchers.
- Versatile Reactive Forms: convenient for self-constructing your desired FRET biomolecules.
- A Complete Set of Dyes: perfectly match your desired fluorescent donors.
- Enhanced Value: competitive price with better performance.
Selection of FRET Donors and Acceptors
FRET can be detected either by the appearance of sensitized fluorescence of the acceptor, by the intensity ratio change of donor/acceptor (if the acceptor is fluorescent), or by the fluorescence decrease of the donor. In the later case, acceptor can be either fluorescent or non-fluorescent. Table 4.1. summarizes our Tide Quencher™ FRET building blocks designed to develop FRET probes for the demanding applications. Based on our in-house research and experience, it is recommended to use the FRET pairs marked in green. The pairs marked in light blue are OK to use, but less efficient. We have proved that these recommended FRET pairs have demonstrated high sensitivity and low background in our protease and nucleic acid detection assays. There are also other factors that you need to consider besides the FRET efficiency, such as pH, multiplexing and buffer interference, etc.
Table 1. Ordering Info For Tide Quencher Products
Cat# ▲ ▼ | Product Name ▲ ▼ | Unit Size ▲ ▼ |
2170 | Tide Quencher™ 2WS acid [TQ2WS acid] | 25 mg |
2178 | Tide Quencher™ 2WS succinimidyl ester [TQ2WS, SE] | 5 mg |
2179 | Tide Quencher™ 2WS maleimide [TQ2WS maleimide] | 1 mg |
2060 | Tide Quencher™ 4WS acid [TQ4WS acid] | 5 mg |
2061 | Tide Quencher™ 4WS amine [TQ4WS amine] | 1 mg |
2062 | Tide Quencher™ 4 CPG [TQ4 CPG] *500 Å* | 100 mg |
2063 | Tide Quencher™ 4 CPG [TQ4 CPG] *1000 Å* | 100 mg |
2064 | Tide Quencher™ 4WS maleimide [TQ4WS maleimide] | 1 mg |
2067 | Tide Quencher™ 4WS succinimidyl ester [TQ4WS SE] | 1 mg |
2068 | Tide Quencher™ 4WS azide [TQ4WS azide] | 1 mg |