Lumiprobe Corporation是美國一家高品質生物技術公司，專業提供分子生物學研究用的活性熒光染料。從2006年開始，我公司生產并銷售生命科學研究和診斷學應用的優質化學藥品。我們的產品主要有：活性染料(Reactive Dye)和SYBR Green I 染料，用于寡核苷酸合成的亞磷酰胺，點擊化學和其它試劑。大部分產品我們都有庫存，可以隨時發到世界各個地方。
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dsGreen Gel Staining Solution, 10000×
Cyanine5 NHS ester
Cyanine3 NHS ester
Sulfo-Cyanine3 carboxylic acid
Cyanine3 carboxylic acid
Azidobutyric acid NHS ester
Azido dye derivatives, 10 mM DMSO solutions
Cy3 azide, 10 mM in DMSO
Cy3.5 azide, 10 mM in DMSO
Cy5 azide, 10 mM in DMSO
Cy5.5 azide, 10 mM in DMSO
Cy7 azide, 10mM in DMSO
5-FAM azide, 10 mM in DMSO
6-FAM azide, 10 mM in DMSO
5-JOE azide, 10 mM in DMSO
5-ROX azide, 10 mM in DMSO
6-ROX azide, 10 mM in DMSO
5-TAMRA azide, 10 mM in DMSO
Alkyne amidite, 5′-terminal
Alkyne amidite, hydroxyprolinol
Alkyne solid support for oligo synthesis
Pentynoic acid STP ester
熒光激活染料（Catalysts and activators）
Copper(II)-TBTA complex, 10 mM in 55% aq. DMSO
SYBR Green I
Gel staining solution, 10000x
For qPCR, 100x solution
脂類活性染料（Dye activated esters）
Cy3 NHS ester
Cy3.5 NHS ester
Cy5 NHS ester
Cy5.5 NHS ester
2D DIGE熒光染料試劑盒（2D DIGE dye kits）-能與Ettan Dige （GE Healthcare）配合使用，或替代使用。
Alkyne amidite, 5′-terminal
Alkyne amidite, hydroxyprolinol
DMS(O)MT aminolink amidite
定制合成熒光雙標記引物（Custom dual-labeled probes）
Cyanine dyes are molecules containing polymethine bridge between two nitrogen atoms with a delocalized charge:
Due to their structure, cyanines have outstandingly high extinction coefficients often exceeding 100,000 Lmol-1cm-1. Different substituents allow to control properties of the chromophore, such as absorbance wavelength, photostability, and fluorescence. For example, absorbance and fluorescence wavelength can be controlled by a choice of polymethine bridge length: longer cyanines possess higher absorbance and emission wavelengths up to near infrared region.
A number of cyanine dyes have been used for life science applications. A series of thiazole and oxazole dyes have been used as DNA- and protein-binding dyes (like TOTO, YOYO, Stains All and others). But the most popular cyanine dyes for life science research were introduced by Alan Waggoner and colleagues of Carnegie Mellon University in early 1990s. The dyes were a modification of cyanine dye Indocyanine Green (ICG) which was used for angiography since 1970s, and they all contain two indolenine rings flanking polymethyne chain. The dyes were found to exhibit low non-specific binding to biomolecules, and have bright fluorescence owing to their huge extinction coefficients and good quantum yields. Once patented, these molecules are now in public domain after expiration of patents of CMU, and are available for purchase from Lumiprobe for research and commercial use as various reactive derivatives, such as?NHS esters,?maleimides,?azides?for Click chemistry, and other derivatives.
There are two varieties of cyanine dyes: non-sulfonated cyanines, and sulfonated cyanines. For many applications they are interchangeable, because their spectral properties are nearly identical. Both sulfonated and non-sulfonated dyes can be used for the labeling of biomolecules such as DNA and proteins. The difference between the dyes is their solubility: sulfo- dyes are water-soluble, and they do not use of organic co-solvent for the labeling in aqueous environment. They are less prone to aggregation in water. There are cases when one of the type of cyanines is desired (see Sulfonated vs non-sulfonated cyanines section below).
Available non-sulfonated dyes incude Cy3, Cy3.5, Cy5, Cy5.5, Cy7, and Cy7.5. Cy stands for ‘cyanine’, and first digit is number of carbon atoms between indolenine groups. Cy2 which is oxazole derivative rather than indolenin, is an exception from this rule. Suffix .5 is added for benzo-fused cyanines. Variation of the structures allows to change fluorescence properties of the molecules, and to cover most important part ot visible and NIR spectrum with several fluorophores.
Most derivatives of non-sulfonated cyanines (except for hydrochlorides of hydrazides and amines) have low aqueous solubility. When these molecules are used for biomolecule labeling, use of organic co-solvent (5-20% of DMF or DMSO) is necessary for efficient reaction. Cyanine dye should be dissolved in organic solvent first, and added to a solution of biomolecule (protein, peptide, amino-labeled DNA) in appropriate aqueous buffer. When conjugation takes place efficiently, the dye reacts before it precipitates.
Fluorescent properties of non-sulfonated cyanines have little dependence on solvent and surrounding. Absorbance and fluorescence spectra of non-sulfonated cyanine dyes are plotted below.
Sulfonated cyanines include additional sulfo-groups which facilitate dissolution of dye molecules in aqueous phase. Charged sulfonate groups decrease aggregation of dye molecules and heavily labeled conjugates.
Currently available sulfonated cyanines include sulfo-Cy3, sulfo-Cy5, and sulfo-Cy7.
Sulfonated cyanines are highly water soluble. No organic co-solvent is needed to perform labeling with these reagents.
Sulfonated vs non-sulfonated cyanines
Sulfonated and non-sulfonated cyanines exhibit very similar fluorescent properties. However, there are a few differences in labeling protocols that should be noticed. Non-sulfonated cyanines must be dissolved in organic co-solvent (DMF or DMSO) prior to use, and added to a solution of target molecule in aqueous buffers. Recommended volume of co-solvent should be 10% for Cy3, Cy5, Cy7, and 15% for .5 counterparts. Sulfo-Cy reagents can be used in purely aqueous conditions. There is also a difference in purification: when dialysis against water or aqueous buffer is used for purification, sulfo-Cy must be used to achieve efficient removal of unreacted dye material. Reactions with both sulfo- and non-sulfo cyanines can be purified by gel filtration, chromatography (HPLC, FPLC, ion exchange), or electrophoresis.
Sulfonated and non-sulfonated cyanines are interchangeable for the labeling of many classes of targets including:
- soluble proteins, which are tolerant to addition of organic co-solvent
- antibodies (use 5-10% of DMSO/DMF)
- DNA and oligonucleotides
- many small molecules
Conjugates produced with similar sulfo- and non-sulfonated reagents (for example, sulfo-Cy5 and Cy5) are very similar in their fluorescent properties, and can be used with various fluorescence instrumentation.
Sulfonated cyanines must be used for:
- sensitive proteins which are denatured by DMF or DMSO
- protein conjugation when purification is done by dialysis
- nanoparticles in aqueous soluitons
- insoluble or hydrophobic proteins
Non-sulfonated cyanines must be used for:
- reactions in organic media (dichloromethane, acetonitrile)
The following cyanine products are available from Lumiprobe.