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AMCA Fluorescent Dyes


Reactive derivatives of AMCA blue fluorescent dye.

AMCA Reagents

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:

  NHS-AMCA Sulfo-NHS-AMCA
Alternative names Succinimydyl-7-amino-4-methylcoumarin-3-acetic acid,
AMCA NHS Ester, (like AMCA-X SE)
Sulfosuccinimydyl-7-amino-4-methyl- coumarin-3-acetic acid, AMCA Sulfo-NHS Ester
Molecular weight 330.29 432.34
Excitation wavelength 345 to 350nm 345 to 350nm
Emission wavelength 440 to 460nm 440 to 460nm
Extinction coefficient ~19,000/M cm ~19,000/M cm
Purity > 90% by HPLC > 90% by HPLC
Solubility Soluble in DMF, DMSO Soluble in water, DMSO
Storage Desiccated at -20°C, protect from moisture, use fresh solutions Desiccated at -20°C, protect from moisture, use fresh solutions
Reactive groups NHS ester, reacts with primary amines at pH 7.0 to 9.0 Sulfo-NHS ester, reacts with primary amines at pH 7.0 to 9.0

 

  AMCA-Hydrazide
Alternative names Aminomethyl-coumarin hydrazide
Molecular weight 247.25
Excitation wavelength 345 to 350nm
Emission wavelength 440 to 460nm
Extinction coefficient ~19,000/M cm
Purity > 90% by HPLC
Solubility Soluble in DMF
Storage Desiccated at 4°C
Reactive groups Hydrazide, reacts with aldehydes (oxidized sugars), or the amine can be conjugated to carboxyls using EDC.

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


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
33005 NHS-AMCA
Formulation: NHS ester of aminomethylcoumarin acetate
Sufficient For: 1 or 2 uses to label a total of 101mg of IgG using typical conditions
5mg Product Instructions for product #33005 NHS-AMCA MSDS for product #33005 NHS-AMCA Certificate of Analysis for product #33005 NHS-AMCA $125.00

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33010 Sulfo-NHS-AMCA
Formulation: Sulfo-NHS ester of aminomethylcoumarin acetate
Sufficient For: 1 or 2 uses to label a total of 77mg of IgG using typical conditions
5mg Product Instructions for product #33010 Sulfo-NHS-AMCA MSDS for product #33010 Sulfo-NHS-AMCA Certificate of Analysis for product #33010 Sulfo-NHS-AMCA $125.00

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33015 Hydrazide-AMCA
Formulation: Hydrazide-activated aminomethylcoumarin acetate
Sufficient For: 1 or 2 uses to label a total of 101mg of IgG using typical conditions
5mg Product Instructions for product #33015 Hydrazide-AMCA MSDS for product #33015 Hydrazide-AMCA Certificate of Analysis for product #33015 Hydrazide-AMCA $131.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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