Tide Fluor™ 3 succinimidyl ester [TF3 SE]Superior replacement for Cy3

Fluorescent dye NHS esters (or succinimidyl esters) are the most popular tool for conjugating dyes to a peptide, protein, antibody, amino-modified oligonucleotide or nucleic acid. NHS esters react readily with the primary amines (R-NH<sub>2</sub>) of proteins, amine-modified oligonucleotides, and other amine-containing molecules. The resulting dye conjugates are quite stable.

The schematic procedures of coupling reaction of aa-RNA and NHS ester-linked Tide Fluor 3; (a) Schematic illustration of the labeling workflow at U22 position. Residues highlighted on secondary structure of U22-aa-rbA in blue represent the first 13 nucleotides synthesized in the initiation stage, residues highlighted in orange, gree, and red represent 3step in elongation stage. The aminoally atom is represented by red dot at position 22, and TF3 is represented by green star. (b) Illustration of reaction between aminoallyl-modified RNA and NHS ester-linked fluorescent dyes. (c) HPLC profiles for original method efficiency. The spectra in black is original method, in red is unreacted aa-RNA. Source: <b>Optimization of N-hydroxysuccinimide ester coupling with aminoallyl-modified RNA for fluorescent labeling</b> by Mengyang Li, <em>Bioengineered</em>, April 2020.

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Additional ordering information

Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
Quotation	Request
International	See distributors
Shipping	Standard overnight for United States, inquire for international

Physical properties

Molecular weight	555.58
Solvent	DMSO

Spectral properties

Correction Factor (280 nm)	0.179
Extinction coefficient (cm ^-1 M ^-1)	75000¹
Excitation (nm)	554
Emission (nm)	578

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	12171501

Overview SDS Protocol

Molecular weight

555.58

Correction Factor (280 nm)

0.179

Extinction coefficient (cm ^-1 M ^-1)

75000¹

Excitation (nm)

554

Emission (nm)

578

Tide Fluor™ 3 (TF3) family has the spectral properties essentially identical to those of Cy3. Compared to Cy3 probes TF3 family has much stronger fluorescence and higher photostability. Additionally their fluorescence is pH-independent from pH 3 to 11. These characteristics make this new dye family a superior alternative to Cy3. TF3-labeled peptides and nucleotides exhibit much stronger fluorescence and higher photostability than the ones labeled with Cy3. In pairing with our Tide Quencher™ 3 (TQ3), a variety of FRET peptides and nucleotides can be developed for detecting proteases and molecular beacons with enhanced sensitivity and stability.

Calculators

Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of Tide Fluor™ 3 succinimidyl ester [TF3 SE]*Superior replacement for Cy3* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

	0.1 mg	0.5 mg	1 mg	5 mg	10 mg
1 mM	179.992 µL	899.96 µL	1.8 mL	9 mL	17.999 mL
5 mM	35.998 µL	179.992 µL	359.984 µL	1.8 mL	3.6 mL
10 mM	17.999 µL	89.996 µL	179.992 µL	899.96 µL	1.8 mL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
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Spectrum

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Spectral properties

Correction Factor (280 nm)	0.179
Extinction coefficient (cm ^-1 M ^-1)	75000¹
Excitation (nm)	554
Emission (nm)	578

Product Family

Name	Excitation (nm)	Emission (nm)	Extinction coefficient (cm ^-1 M ^-1)	Correction Factor (280 nm)
Tide Fluor™ 1 succinimidyl ester [TF1 SE]Superior replacement for EDANS	341	448	20000	0.187
Tide Fluor™ 2, succinimidyl ester [TF2 SE]Superior replacement for fluorescein	503	525	75000	0.09
Tide Fluor™ 5WS succinimidyl ester [TF5WS SE]Superior replacement for Cy5	649	664	250000	0.027
Tide Fluor™ 4, succinimidyl ester [TF4 SE]Superior replacement for ROX and Texas Red	578	602	90000	0.436
Tide Fluor™ 6WS succinimidyl ester [TF6WS SE]Superior replacement for Cy5.5	682	701	220000	0.101
Tide Fluor™ 7WS, succinimidyl ester [TF7WS SE]Superior replacement for Cy7	756	780	275000	0.049
Tide Fluor™ 8WS, succinimidyl ester [TF8WS SE]Near Infrared Emission	785	801	250000	0.109
Tide Fluor™ 3WS succinimidyl ester [TF3WS SE] Superior replacement for Cy3	551	563	150000	0.079
Tide Fluor™ 2WS succinimidyl ester [TF2WS SE] Superior replacement for FITC	491	516	75000	0.11
Tide Quencher™ 3 succinimidyl ester [TQ3 SE]	-	-	22000	0.091
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Images

Figure 1. Fluorescent dye NHS esters (or succinimidyl esters) are the most popular tool for conjugating dyes to a peptide, protein, antibody, amino-modified oligonucleotide or nucleic acid. NHS esters react readily with the primary amines (R-NH₂) of proteins, amine-modified oligonucleotides, and other amine-containing molecules. The resulting dye conjugates are quite stable.

Figure 2. The schematic procedures of coupling reaction of aa-RNA and NHS ester-linked Tide Fluor 3; (a) Schematic illustration of the labeling workflow at U22 position. Residues highlighted on secondary structure of U22-aa-rbA in blue represent the first 13 nucleotides synthesized in the initiation stage, residues highlighted in orange, gree, and red represent 3step in elongation stage. The aminoally atom is represented by red dot at position 22, and TF3 is represented by green star. (b) Illustration of reaction between aminoallyl-modified RNA and NHS ester-linked fluorescent dyes. (c) HPLC profiles for original method efficiency. The spectra in black is original method, in red is unreacted aa-RNA. Source: Optimization of N-hydroxysuccinimide ester coupling with aminoallyl-modified RNA for fluorescent labeling by Mengyang Li, Bioengineered, April 2020.

Citations

View all 9 citations: Citation Explorer

Optimization of N-hydroxysuccinimide ester coupling with aminoallyl-modified RNA for fluorescent labeling
Authors: Li, Mengyang
Journal: Bioengineered (2020): 599--606

To what extent do fluorophores bias the biological activity of peptides? A practical approach using membrane-active peptides as models
Authors: Cavaco, Marco and P{\'e}rez-Peinado, Clara and Valle, Javier and Silva, R{\'u}ben DM and Correia, Jo{\~a}o DG and Andreu, David and Castanho, Miguel ARB and Neves, Vera
Journal: Frontiers in bioengineering and biotechnology (2020): 552035

A mechanistic model to predict effects of cathepsin B and cystatin C on β-amyloid aggregation and degradation
Authors: Perlenfein, Tyler J and Murphy, Regina M
Journal: Journal of Biological Chemistry (2017): jbc--M117

Real-Time Detection of a Self-Replicating RNA Enzyme
Authors: Olea, Charles and Joyce, Gerald F
Journal: Molecules (2016): 1310

Maternal serum glycosylated fibronectin as a point-of-care biomarker for assessment of preeclampsia
Authors: Rasanen, Juha and Quinn, Matthew J and Laurie, Amber and Bean, Eric and Roberts, Charles T and Nagalla, Srinivasa R and Gravett, Michael G
Journal: American journal of obstetrics and gynecology (2015): 82--e1

Development of Multi-Parametric/Multimodal Spectroscopy Apparatus for Characterization of Functional Interfaces
Authors: Zhou, Lang and Arugula, Mary and Easley, Christopher J and Shannon, Curtis and Simonian, Aleks and r, undefined
Journal: ECS Transactions (2015): 9--16

Array of biodegradable microrafts for isolation and implantation of living, adherent cells
Authors: Wang, Yuli and Phillips, Colleen N and Herrera, Gabriela S and Sims, Christopher E and Yeh, Jen Jen and Allbritton, Nancy L
Journal: RSC advances (2013): 9264--9272

Development of SNAP-Tag Fluorogenic Probes for Wash-Free Fluorescence Imaging
Authors: Sun, Xiaoli and Zhang, Aihua and Baker, Brenda and Sun, Luo and Howard, Angela and Buswell, John and Maurel, Damien and Masharina, Anastasiya and Johnsson, Kai and Noren, Christopher J and others, undefined
Journal: ChemBioChem (2011): 2217--2226

FERRAMENTAS PARA ESTUDO DA BIOLOGIA DE GPCRS (G-PROTEIN COUPLED RECEPTORS)
Authors: Soriani, Frederico Marianetti and Russo, Remo Castro

References

View all 25 references: Citation Explorer

Time-resolved FRET method for typing polymorphic alleles of the human leukocyte antigen system by using a single DNA probe
Authors: Andreoni A, Bondani M, Nardo L.
Journal: Photochem Photobiol Sci (2009): 1202

Tumor-specific detection of an optically targeted antibody combined with a quencher-conjugated neutravidin "quencher-chaser": a dual "quench and chase" strategy to improve target to nontarget ratios for molecular imaging of cancer
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Journal: Bioconjug Chem (2009): 147

The detection of platelet derived growth factor using decoupling of quencher-oligonucleotide from aptamer/quantum dot bioconjugates
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Journal: Nanotechnology (2009): 175503

Development of a cell-based hepatitis C virus infection fluorescent resonance energy transfer assay for high-throughput antiviral compound screening
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An improved cell-penetrating, caspase-activatable, near-infrared fluorescent peptide for apoptosis imaging
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Journal: Bioconjug Chem (2009): 702

Feasibility of single nucleotide polymorphism genotyping with a single-probe by time-resolved Forster resonance energy transfer
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Journal: Mol Cell Probes (2009): 119

Photodynamic molecular beacon triggered by fibroblast activation protein on cancer-associated fibroblasts for diagnosis and treatment of epithelial cancers
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Rapid detection and quantification of Propionibacteriaceae
Authors: Goldschmidt P, Ferreira CC, Degorge S, Benallaoua D, Boutboul S, Laroche L, Batellier L, Chaumeil C.
Journal: Br J Ophthalmol (2009): 258

Evaluation of tetramethylrhodamine and black hole quencher 1 labeled probes and five commercial amplification mixes in TaqMan real-time RT-PCR assays for respiratory pathogens
Authors: Yang GP, Erdman DD, Tondella ML, Fields BS.
Journal: J Virol Methods (2009): 288

Design of FRET-TaqMan probes for multiplex real-time PCR using an internal positive control
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Journal: Biotechniques (2009): 519