与氨基反应的荧光探针

Highlights of Rhodamine and TRITC Labeling Kits:

∙ ∙ ∙ ∙ ∙

Fast – label and purify antibody in about one hour

Amine-specific labeling – the NHS-ester or isothiocyanate efficiently labels antibodies and other purified proteins at primary amines (lysine side chains) Optimized procedure – following the standard protocol results in antibodies with excellent dye:protein ratios for optimum activity and fluorescence

Single-use fluors – no need to weigh tiny amounts powder; kits contain single-use vials of reagent

Efficient purification – kits include purification resin and easy-to-use spin columns, ensuring rapid and efficient removal of non-reacted dye and excellent protein recovery

Applications for Rhodamine Labeling Reagents:

∙ ∙ ∙

Label antibodies for use as immunofluorescent probes Label oligonucleotides for hybridization probes Detect proteins in gels and on Western blots

Properties of

Rhodamine Dyes:

Thermo Scientific Pierce Rhodamine Dyes are mixtures of isomers with reactive groups attached at the 5- and 6-positions of the bottom ring. The properties of these isomers are indistinguishable in terms of excitation and emission spectra, and for protein applications there is no need to isolate a specific isomer.

TRITC is the base tetramethylrhodamine molecule functionalized with an isothiocyanate reactive group (–N=C=S) at one of two hydrogen atoms on the bottom ring of the structure. This derivative is reactive towards primary amine groups on proteins, peptides and other biomolecules. NHS-Rhodamine is activated with the N-hydroxy-succinimidyl-ester (NHS ester) functional group. Compared to TRITC, the NHS-ester deriviative has greater specificity toward primary amines in the presence of other nucleophiles and results in a more stable linkage following labeling. Texas Red* Sulfonyl Chloride is a long-wavelength derivative of rhodamine that is modified with sulfonyl chloride for reaction to primary amines. We recommend

DyLight

594 NHS Ester as an alternative to Texas Red Sulfonyl Chloride because it is brighter, more photostable and has a more specific labeling chemistry that can be used at physiologic pH. AMCA (aminomethylcoumarin acetate) is a blue fluorescent dye whose reactive derivatives are used as contrasting probes for double and triple labeling in immunofluorescence microscopy, arrays and in situ hybridization. The desirable properties of AMCA dyes also include a relatively large Stoke's shift and resistance to photobleaching. NHS-AMCA and Sulfo-NHS-AMCA are reactive towards primary amine groups on proteins, peptides and other biomolecules, while AMCA HPDP reacts with sulfhydryls, and AMCA hydrazide is used to label glycosylation sites. AMCA Hydrazide can also be reacted with carboxyl groups using the crosslinker EDC. AMCA has an excitation maximum at 345nm and an emission maximum at 450nm.

与巯基反应的荧光探针 Properties of AMCA Dyes:

与氨基反应的荧光探针

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