AEC Substrate Kit (ab64252)
Product nameAEC Substrate Kit
Tested applicationsSuitable for: IHC-Pmore details
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.
Storage instructionsStore at +4°C.
Storage bufferpH: 4.7
Constituents: 0.03% Hydrogen peroxide, 0.1% 3-amino-9-ethyl carbazole, 3% Dimethyl formamide
Concentration information loading...
RelevanceAEC (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.
- 3 amino 9 ethylcarbazole
The Abpromise guarantee
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.
Use at an assay dependent dilution.
Use at an assay dependent dilution.
Immunostaining using anti-hMMP-9 murine monoclonal antibody.Slides of superficial spread melanoma tumors were incubated with the hMMP-9 antibody (ab58803, 1:300). Alkanie phosphatas esecondary antibody was stained with AEC (3-Amino-9-EthylCarbazole, ab 64252, Abcam, Cambridge) for 15 min and slides were counterstained with hematoxylin for examination.
Immunostaining for TGM2 and SOX2 staining in GSC 158 and GSC 157 cells.(a) Positive TGM2 and negative SOX2 staining in GSC 158 cells. (b) Negative TGM2 and positive SOX2 staining in GSC 157 cells. Cells were incubated with primary monoclonal Abs for 1 h at +37 °C (for SOX2, ab6734, 1/200) following the Lab Vision Ready-To-Use AEC Substrate System (ab64252) according to the manufacturer's instructions.
Human colon carcinoma stained with anti-Keratin 20 antibody using AEC Substrate System (Ready to Use) (ab64252).
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
ab64252 has been referenced in 13 publications.
- Sforna M et al. Characterization of Primary Cultures of Normal and Neoplastic Canine Melanocytes. Animals (Basel) 11:N/A (2021). PubMed: 33802040
- Silvestri S et al. E-Cadherin Expression in Canine Melanocytic Tumors: Histological, Immunohistochemical, and Survival Analysis. Vet Pathol 57:608-619 (2020). PubMed: 32578507
- Gualandi M et al. Oncogenic ALKF1174L drives tumorigenesis in cutaneous squamous cell carcinoma. Life Sci Alliance 3:N/A (2020). PubMed: 32312912
- Trus I et al. Zika Virus with Increased CpG Dinucleotide Frequencies Shows Oncolytic Activity in Glioblastoma Stem Cells. Viruses 12:N/A (2020). PubMed: 32466170
- Yura Y et al. Endothelial cell-specific redox gene modulation inhibits angiogenesis but promotes B16F0 tumor growth in mice. FASEB J 33:14147-14158 (2019). PubMed: 31647879