Aec 3 amino 9 ethyl carbazole chromogen

Manufactured by Thermo Fisher Scientific

Sourced in United States

AEC (3-amino-9-ethyl carbazole) is a chromogenic substrate used in immunohistochemistry and enzyme-linked immunosorbent assay (ELISA) applications. It produces a reddish-brown colored precipitate when oxidized by peroxidase enzymes, allowing for the visualization and detection of target analytes.

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Lab products found in correlation

Ultravision lp detection system, by Thermo Fisher Scientific (4 mentions) Peroxidase blocking, by Agilent Technologies (4 mentions) Ki 67 antibody, by Agilent Technologies (2 mentions) Ki 67 antibody, by Cell Signaling Technology (1 mentions) Schiff reagent, by Merck Group (1 mentions) R script, by R Project for Statistical Computing (1 mentions) Halo image analysis platform, by Indica Labs (1 mentions) Ucp1 antibody, by Abcam (1 mentions)

4 protocols using aec 3 amino 9 ethyl carbazole chromogen

Automated Quantification of Adipocyte and Apoptosis

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Immunohistochemical stainings of formalin-fixed, paraffin embedded tissues were performed after antigen retrieval (93 °C, 15 min at pH 6) and peroxidase blocking (Agilent, Foster City, CA, United States) using the UltraVision LP detection system (Thermo Fisher Scientific, Waltham, MA, United States) according to the manual for cleaved caspase 3 antibody (9661, 1:50, Cell Signalling Technology, Denver, MA, USA) or Ki67 antibody (12202, 1:400, Cell Signalling Technology, Denver, MA, USA).
AEC (3-amino-9-ethyl carbazole) chromogen (Thermo Fisher Scientific, Waltham, MA, United States) was used for colour detection. Counterstaining with hematoxylin was done on all slides.
White adipocyte cell size was quantified by using an automated plugin for ImageJ (Adiposoft^{99 (link)}) and quantification of crown-like structures and cleaved caspase 3 positive cells was performed by using ImageJ. Liver lipid droplets in H&E sections were counted using VisioPharm version 2019.09.

Reinisch I., Michenthaler H., Sulaj A., Moyschewitz E., Krstic J., Galhuber M., Xu R., Riahi Z., Wang T., Vujic N., Amor M., Zenezini Chiozzi R., Wabitsch M., Kolb D., Georgiadi A., Glawitsch L., Heitzer E., Schulz T.J., Schupp M., Sun W., Dong H., Ghosh A., Hoffmann A., Kratky D., Hinte L.C., von Meyenn F., Heck A.J., Blüher M., Herzig S., Wolfrum C, & Prokesch A. (2024). Adipocyte p53 coordinates the response to intermittent fasting by regulating adipose tissue immune cell landscape. Nature Communications, 15, 1391.

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Multimodal Liver Tissue Analysis

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For Oil Red O staining, 6-µm thin cryosections were mounted on poly-l-lysine-coated slides and dried overnight. Working solution of Oil Red O was prepared by mixing stock solution [0.5% (w/v) in 2-propanol] 3:2 with water followed by filtration. Working solution was incubated for 10 min. To stain liver glycogen, formalin-fixed, paraffin-embedded livers were cleared and incubated in 1% periodic acid (25 min) followed by Schiff reagent (Sigma-Aldrich, St. Louis, MO, USA; 25 min) and rinsing in SO₂ water. Immunohistochemical staining of formalin-fixed, paraffin-embedded livers was performed after antigen retrieval (120°C, 7 min at pH 9) and peroxidase blocking (Dako, Glostrup, Denmark) using the UltraVision LP detection system (Thermo Fisher Scientific) according to the manual with 1 ng/µl Ki-67 antibody (M728; Dako). For color reaction, AEC (3-amino-9-ethylcarbazole) chromogen (Thermo Fisher Scientific) was used. Stainings were performed in a LabVision 2D autostainer (Thermo Fisher Scientific). Mouse IgG₁ was used as negative control. Counterstaining with hematoxylin was done on all slides. All quantifications were performed by Ilastik Interactive Learning and Segmentation software (http://ilastik.org/). Automatic counting of segments was done by a customized R script (R Foundation for Statistical Computing, Vienna, Austria; http://www.r-project.org/).

Prokesch A., Graef F.A., Madl T., Kahlhofer J., Heidenreich S., Schumann A., Moyschewitz E., Pristoynik P., Blaschitz A., Knauer M., Muenzner M., Bogner-Strauss J.G., Dohr G., Schulz T.J, & Schupp M. (2016). Liver p53 is stabilized upon starvation and required for amino acid catabolism and gluconeogenesis. The FASEB Journal, 31(2), 732-742.

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Immunohistochemical Analysis of Xenograft Samples

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Immunohistochemical staining of formalin-fixed, paraffin-embedded xenografts was performed after antigen retrieval (120°C, 7 min at pH 9) and peroxidase blocking (Dako) using the UltraVision LP detection system (Thermo Fisher Scientific) according to the manual with Ki-67 antibody (1 ng/ml; M728; Dako). For color reaction, AEC (3-amino-9-ethyl carbazole) chromogen (Thermo Fisher Scientific) was used. Counterstaining with hematoxylin was done on all slides. Quantitative analysis was performed by Halo image analysis platform (Indica Labs) using random forest tissue classifier. Picrosirius red staining was used to assess cirrhosis in HCC nodules of DEN-treated mice.

Krstic J., Reinisch I., Schindlmaier K., Galhuber M., Riahi Z., Berger N., Kupper N., Moyschewitz E., Auer M., Michenthaler H., Nössing C., Depaoli M.R., Ramadani-Muja J., Usluer S., Stryeck S., Pichler M., Rinner B., Deutsch A.J., Reinisch A., Madl T., Chiozzi R.Z., Heck A.J., Huch M., Malli R, & Prokesch A. (2022). Fasting improves therapeutic response in hepatocellular carcinoma through p53-dependent metabolic synergism. Science Advances, 8(3), eabh2635.

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Immunohistochemistry of Brown Adipose Tissue

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Immunohistochemical staining of formalin-fixed, paraffin-embedded BAT depots was performed after antigen retrieval (93°C, 15 min at pH 6) and peroxidase blocking (Agilent, Foster City, CA, United States, S202386-2) using the UltraVision LP detection system (Thermo Fisher Scientific, Waltham, MA, United States, 12643997) according to the manual with UCP1-antibody (1.25 µg/ml; Abcam, MA, United Kingdom, 10983). AEC (3-amino-9-ethyl carbazole) chromogen (Thermo Fisher Scientific, Waltham, MA, United States, 001122) was used for color detection. Counterstaining with hematoxylin was done on all slides. Hematoxylin and eosin stainings were quantified by using ImageJ Brown adipocyte area was indicated as square pixels.

Reinisch I., Klymiuk I., Michenthaler H., Moyschewitz E., Galhuber M., Krstic J., Domingo M., Zhang F., Karbiener M., Vujić N., Kratky D., Schreiber R., Schupp M., Lenihan-Geels G., Schulz T.J., Malli R., Madl T, & Prokesch A. (2022). p53 Regulates a miRNA-Fructose Transporter Axis in Brown Adipose Tissue Under Fasting. Frontiers in Genetics, 13, 913030.

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