Overview

  • Product name

    AEC Substrate Kit
  • Description

    AEC Substrate Kit
  • Tested applications

    Suitable for: IHC-Pmore details
  • General notes

    AEC substrate kit for use in IHC using HRP / peroxidase detection. Ready to use formulation.

    Previously called AEC Substrate System (Ready to Use).


    IHC protocol suitable for use with AEC Substrate Kit ab64252:

    1. Deparaffinize and rehydrate formalin-fixed paraffin-embedded tissue section.

    2. Use appropriate antigen retrieval buffer or enzyme (primary antibody dependent) to treat sections. Wash 3 times in buffer.

    3. Add enough hydrogen peroxide blocking solution to cover the sections. Incubate for 10 minutes. Wash 2 times in buffer. If necessary, use avidin biotin blocking.

    4. Apply protein block (or normal serum from same species as secondary antibody) and incubate for 5 minutes at room temperature to block nonspecific background staining. Wash once in buffer.

    5. Apply primary antibody in antibody diluent and incubate.

    6. Wash 4 times in buffer. Incubate slide with biotinylated secondary antibody (or HRP polymer secondary antibody and skip step 7). Wash 4 times in buffer.

    7. Apply streptavidin-HRP and incubate for 10 minutes at room temperature.

    8. Rinse 4 times in buffer. Place slide in DAB substrate or AEC Substrate ab64252 and incubate until desired color is achieved (1-10 mins, recommended 10 mins for AEC). Rinse 4 times in buffer.

    9. Add enough drops of hematoxylin to cover the section. Incubate for 1 minute.

    10. Rinse 7-8 times in tap water. Add mounting medium to cover the section.


    Find complete IHC kits, and reagents for antigen retrieval, blocking, signal amplification, visualization, counterstaining, and mounting in the IHC kits and reagents guide.

Properties

  • Form

    Liquid
  • Storage instructions

    Store at +4°C.
  • Storage buffer

    Constituents: 0.03% Hydrogen peroxide, 3% DMF

    stabilizers
  • Concentration information loading...
  • Research areas

  • Relevance

    AEC (3-Amino-9-Ethylcarbazole) is a widely used chromogen for immunohistochemical staining. AEC produces a red end product that is soluble in alcohol and must be used with an aqueous counterstain and mounting media. AEC produces a clean, very intense stain that is free of unwanted background.
  • Alternative names

    • 3 amino 9 ethylcarbazole

Applications

Our Abpromise guarantee covers the use of ab64252 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P Use at an assay dependent dilution.

Images

  • Human colon carcinoma stained with anti-Keratin 20 antibody using AEC Substrate System (Ready to Use) (ab64252).

References

This product has been referenced in:

  • Maliniemi P  et al. Biological and clinical significance of tryptophan-catabolizing enzymes in cutaneous T-cell lymphomas. Oncoimmunology 6:e1273310 (2017). Read more (PubMed: 28405495) »
  • Brachelente C  et al. Transcriptome Analysis of Canine Cutaneous Melanoma and Melanocytoma Reveals a Modulation of Genes Regulating Extracellular Matrix Metabolism and Cell Cycle. Sci Rep 7:6386 (2017). Read more (PubMed: 28743863) »
See all 3 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer

Thank you for contacting us.
The kit will work with the rabbit and mouse primary antibodies.
You can try citrate buffer pH for a start and Tris/EDTA in case the staining does not work. Please check the following for protocol and troubleshooting of;
https://www.abcam.com/index.html?pageconfig=resource&rid=11384
https://www.abcam.com/index.html?pageconfig=resource&rid=11467
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

Read More

Answer

Thank you for contacting us.
Heat mediated antigen retrieval method with microwave or pressure cooker is recommended for this ab.
The suitable detection system depends on the substrate you want to use. e.g. with DAB or AEC, we have following products;
https://www.abcam.com/DAB-Enhancer-ab675.html
https://www.abcam.com/DAB-Substrate-Kit-ab64238.html
https://www.abcam.com/AEC-Substrate-System-Ready-to-Use-ab64252.html
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

Read More

Answer


Yes, Mouse and Rabbit Specific HRP/DAB (ABC) detection IHC kit (ab64264) and AEC Substrate System (Ready to Use) (ab64252) can be used with rat sections.

Read More

Answer

Thank you very much for your enquiry. We do not currently sell substrates such as AP and HRP that have been tested in ELISPOT. This doesn’t mean that these products are not suitable, we just have not tested them in this application. The 4 products identified (ab7413, ab7468, ab64252, ab103742) have been tested in IHC and ab7413 has also been tested in dot blots and WB. This may be a more suitable one to try but unfortunately we cannot guarantee that it would be suitable. I would consider reviewing what ELISPOT kits we have available as we may have one that is suitable. I would be happy to offer any further advice and hope this information helps.

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