Thermo Scientific AMCA Reagents are derivatives of AMCA blue fluorescent dye that are activated for easy and reliable labeling of antibodies, proteins and other molecules for use as fluorescent probes.
AMCA (aminomethylcoumarin acetate) is a blue fluorescent dye with reactive derivatives that are used as contrasting probes for double- and triple-labeling in immunofluorescence microscopy, arrays and in situ hybridization. The desirable properties of AMCA dyes 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. AMCA-Hydrazide is used to label glycosylation sites or for conjugation to carboxyl groups using the crosslinker EDC. AMCA has an excitation maximum at 345nm and an emission maximum at 450nm.
Highlights:
- AMCA – activated derivatives of blue-fluorescent aminomethylcoumarin acetate dye
- Amine-specific labeling – NHS-ester and Sulfo-NHS-ester varieties of AMCA efficiently label antibodies and other purified proteins at primary amines (lysine side chains)
- Glycoprotein labeling – hydrazide variety of AMCA labels oxidized sugar groups in glycoproteins or other carbohydrates
Properties of AMCA Dyes:
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NHS-AMCA
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Sulfo-NHS-AMCA
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Alternative names
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Succinimydyl-7-amino-4-methylcoumarin-3-acetic acid,
AMCA NHS Ester, (like AMCA-X SE)
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Sulfosuccinimydyl-7-amino-4-methyl- coumarin-3-acetic acid, AMCA Sulfo-NHS Ester
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Molecular weight
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330.29
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432.34
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Excitation wavelength
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345 to 350nm
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345 to 350nm
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Emission wavelength
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440 to 460nm
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440 to 460nm
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Extinction coefficient
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~19,000/M cm
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~19,000/M cm
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Purity
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> 90% by HPLC
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> 90% by HPLC
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Solubility
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Soluble in DMF, DMSO
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Soluble in water, DMSO
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Storage
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Desiccated at -20°C, protect from moisture, use fresh solutions
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Desiccated at -20°C, protect from moisture,
use fresh solutions
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Reactive groups
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NHS ester, reacts with primary amines at pH 7.0 to 9.0
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Sulfo-NHS ester, reacts with primary amines at pH 7.0 to 9.0
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AMCA-Hydrazide
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Alternative names
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Aminomethyl-coumarin hydrazide
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Molecular weight
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247.25
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Excitation wavelength
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345 to 350nm
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Emission wavelength
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440 to 460nm
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Extinction coefficient
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~19,000/M cm
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Purity
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> 90% by HPLC
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Solubility
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Soluble in DMF
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Storage
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Desiccated at 4°C
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Reactive groups
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Hydrazide, reacts with aldehydes (oxidized sugars), or the amine can be conjugated to carboxyls using EDC.
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Related Resources:
| DyLight 350 and DyLight 405 are newer derivatives of AMCA dye, tailored for various chemical and biological applications where greater photostability and fluorescence intensity, pH independence, and a narrower emission spectrum are required. |
Review of Fluorescent Probes
Tech Tip #31 – Calculate dye:protein (F/P) molar ratios
Related Products:
DyLight Fluor Labeling Reagents and Kits
Fluorescent Labeling – Top-level menu of all reagents
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