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3-Amino-9-ethylcarbazole

(Synonyms: AEC) 目录号 : GC25016

3-Amino-9-ethylcarbazole (AEC) is a chemical compound commonly used as a chromogenic substrate in immunohistochemistry, specifically for visualizing sections stained with HRP-conjugated secondary antibodies.

3-Amino-9-ethylcarbazole Chemical Structure

Cas No.:132-32-1

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产品描述

3-Amino-9-ethylcarbazole (AEC) is a chemical compound commonly used as a chromogenic substrate in immunohistochemistry, specifically for visualizing sections stained with HRP-conjugated secondary antibodies.

[1] Stack EC, et al. Methods. 2014 Nov;70(1):46-58.

Chemical Properties

Cas No. 132-32-1 SDF Download SDF
别名 AEC
分子式 C14H14N2 分子量 210.27
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Research Update

Metachromasia of 3-Amino-9-ethylcarbazole (AEC) and its prevention in immunoperoxidase techniques

Histochemistry 1987;86(5):471-8.PMID:2438258DOI:10.1007/BF00500619.

3-Amino-9-ethylcarbazole (AEC) used as chromogen in immunoperoxidase techniques normally has an intense, red colour. However, as an inconstant phenomenon, a pale yellowish-green reaction product severely impairing the evaluation can be observed. In order to circumvent this undesired effect, factors such as tissue fixative, proteolytic digestion, antibody concentrations and incubation time of the primary antibody were analyzed. The most important factor inducing a change in colour is probably the inadequately high local peroxidase concentration arising as the consequence of high amounts of bound primary antibody. This high enzyme concentration might cause metachromasia of AEC by producing the yellowish-green quinone-di-imine form of the substrate. As could be shown by spectrophotometry in test tube experiments, AEC metachromasia was proven to be enzyme dependent. Thus, the best way to trigger the local enzyme concentration on a tissue section to adequate levels appears to be the dilution of the primary antibody.

A selective optical sensor for picric acid assay based on photopolymerization of 3-(N-methacryloyl) amino-9-ethylcarbazole

Anal Chim Acta 2006 Jun 16;570(2):170-5.PMID:17723396DOI:10.1016/j.aca.2006.04.026.

A novel optical sensor based on covalent immobilization for picric acid assay has been described. To improve the stability of the sensor, a terminal double bond was attached to the fluorescent compound, 3-Amino-9-ethylcarbazole (AEC), via methacryloyl chloride. The resultant compound, 3-(N-methacryloyl) amino-9-ethylcarbazole (MAEC) was copolymerized with 2-hydroxypropyl methacrylate on surface-modified quartz glass plates by UV irradiation. The resulting optical sensor (optode membrane) was used to determine picric acid based on fluorescence quenching. It shows a linear response toward picric acid in the concentration range of 9.33 x 10(-8) to 9.33 x 10(-5) mol l(-1), with rapid response, high stability and good selectivity to picric acid.

A novel fluorescence sensor based on covalent immobilization of 3-Amino-9-ethylcarbazole by using silver nanoparticles as bridges and carriers

Anal Chim Acta 2009 Mar 23;636(2):205-9.PMID:19264169DOI:10.1016/j.aca.2009.01.034.

A novel technique of covalent immobilization of indicator dyes in the preparation of fluorescence sensors is developed. Silver nanoparticles are used as bridges and carriers for anchoring indicator dyes. 3-Amino-9-ethylcarbazole (AEC) was employed as an example of indicator dyes with terminal amino groups and covalently immobilized onto the outmost surface of a quartz glass slide. First, the glass slide was functionalized by (3-mercaptopropyl) trimethoxysilane (MPS) to form a thiol-terminated self-assembled monolayer, where silver nanoparticles were strongly bound to the surface through covalent bonding. Then, 16-mercaptohexadecanoic acid (MHDA) was self-assembled to bring carboxylic groups onto the surface of silver nanoparticles. A further activation by using 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) converted the carboxylic groups into succinimide esters. Finally, the active succinimide esters on the surface of silver nanoparticles were reacted with AEC. Thus, AEC was covalently bound to the glass slide and an AEC-immobilized sensor was obtained. The sensor exhibited very satisfactory reproducibility and reversibility, rapid response and no dye-leaching. Rutin can quench the fluorescence intensity of the sensor and be measured by using the sensor. The linear response of the sensor to rutin covers the range from 2.0 x 10(-6) to 1.5 x 10(-4) molL(-1) with a detection limit of 8.0 x 10(-7) molL(-1). The proposed technique may be feasible to the covalent immobilization of other dyes with primary amino groups.

Novel chromogenic substrates with metal chelating properties for the histochemical detection of peroxidasic activity, derived from 3-Amino-9-ethylcarbazole (AEC) and 3,6-diamino-9-ethylcarbazole

Cell Mol Biol (Noisy-le-grand) 2000 Nov;46(7):1191-212.PMID:11075949doi

For staining of peroxidase activity routinely employed 3-Amino-9-ethylcarbazole 1 (AEC) was chemically modified in order to obtain chromogenic enzyme substrates with improved staining properties. In conclusion of systematically structure/staining considerations of a series of novel substrates, it can be generalized that the performance of traditional chromogenic peroxidase amine-substrates is accessible an considerably improvement in terms of sensitivity and adaptibility for various application purposes (solubility and color of the reaction product, electron dense and osmiophilic properties, ...) by attachment of chelating N-benzyl-moieties making available highly efficient the well known metal catalytic effect in a proposed intramolecular way. Thus, the model compounds 3(arylmethyl)amino-9-ethyl-carbazole 4 and 3,6-bis-(arylmethyl)amino-9-ethyl-carbazole 7 were synthesized by condensation of 3-Amino-9-ethylcarbazole 1 (AEC) or the corresponding 3,6-diamine 5 with aromatic aldehydes 2. The resulting Schiff bases 3 and 6 were subsequently reduced with sodium borohydride. The obtained benzylamines 4 and 7 were examined as chromogenic substrates: 1) qualitatively in test tube experiments concerning color, precipitation behavior and solubility of the precipitates, 2) quantitatively by means of electroblotted dilution series of horseradish peroxidase, and finally in a biological environment for the localization of endogenous peroxidasic activity 3) in native cryotome tissues of Wistar rats. 4) The usefulness of the new approach for electron microscopy was revealed, too. Thus the discrimination between internum and externum of specific granules after osmium tetroxide contrastate was higher if compared with results obtained by the Karnovsky protocol. The wide spread variation of substitution patterns of the novel reagents gave reason for structure-reactivity considerations and ongoing leading structures. The stereochemical and electronic factors as well as competing reaction pathways governing the reaction course are briefly discussed. In addition, the metal associating reagents are highly effective in oxidative side-coupling reactions with aromatic amine or phenol-additives exemplified here by means of 4-amino-N,N-diphenylamine. The reagents 4 and 7 are obtainable in a simple in situ synthesis, too, offering in principle a 'chemical construction unit'. The demonstrated approach is of general interest for bioanalytical applications offering an access to potentially novel chromogens and electron opaque markers for the detection of peroxidasic activity/hydroperoxides or related redox enzyme systems.

Bioassay of 3-Amino-9-ethylcarbazole hydrochloride for possible carcinogenicity

Natl Cancer Inst Carcinog Tech Rep Ser 1978;93:1-193.PMID:12806398doi

A bioassay of 3-Amino-9-ethylcarbazole (hydrochloride) for possible carcinogenicity was conducted by administering the test chemical in feed to Fischer 344 rats and B6C3F1 mice. Both the free amine and the hydrochloride salt were used. Groups of 50 rats of each sex and 50 mice of each sex were administered the test chemical at one of two doses, either 800 or 2,000 ppm for rats and either 800 or 1,200 ppm for mice, for 78 weeks. The rats were then observed for an additional 26-29 weeks, and the mice for an additional 16-17 weeks. Controls consisted of groups of 50 untreated rats of each sex and 50 untreated mice of each sex; separate controls were used for the groups of animals administered the different doses. All surviving rats were killed at 104-110 weeks; all surviving mice were killed at 94-97 weeks. Since the suppliers of the low-dose rats and mice differed from those of the corresponding low-dose controls, while the suppliers for the high-dose rats and mice were the same as those of the corresponding high-dose controls, comparisons of the high-dose groups with their corresponding controls were the most appropriate. Furthermore, since the low-dose animals did not receive the same regimen of administration of the test compound as that received by the high-dose animals, and since tests using the low-dose groups were not performed concurrently with those using the high-dose groups, analyses of the dose-related trends were not possible. Although the interpretation of results of the study was based primarily on comparisons of high-dose groups with their respective controls, the results obtained with the low-dose groups, regardless of the indicated complicating factors, supported the interpretation. Neoplasms of the liver were observed in significant incidences in rats and mice of both sexes. In male rats, hepatocellular carcinomas alone were significantly higher (P3-Amino-9-ethylcarbazole (hydrochloride) was carcinogenic for the liver, inducing hepatocellular carcinomas in Fischer 344 rats and B6C3F1 mice of both sexes. Other tumors induced in the rats were carcinomas or papillomas of the integumentary system in males, carcinomas of the Zymbal's gland of the ear in males and females, and adenocarcinomas of the uterus.