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    aec-substratekit-ab64252.pdf

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AEC Substrate Kit (ab64252)

  • Datasheet
  • SDS
Submit a review Q&A (4)References (13)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - AEC Substrate Kit (ab64252)
  • Immunohistochemistry (Frozen sections) - AEC Substrate Kit (ab64252)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - AEC Substrate System (Ready to Use) (ab64252)

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Overview

  • Product name

    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

    pH: 4.7
    Constituents: 0.03% Hydrogen peroxide, 0.1% 3-amino-9-ethyl carbazole, 3% Dimethyl formamide

    stabilizers
  • Concentration information loading...
  • Research areas

    • Kits/ Lysates/ Other
    • Tools and Reagents
    • IHC Tools/ Reagents
    • Kits/ Lysates/ Other
    • Kits
    • IHC tools
    • Substrate
  • 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

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.

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

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - AEC Substrate Kit (ab64252)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - AEC Substrate Kit (ab64252)Image from Kryza et al. PLoS One; 11(2) :e0149387. doi: 10.1371/journal.pone.0149387. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/
    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.
  • Immunohistochemistry (Frozen sections) - AEC Substrate Kit (ab64252)
    Immunohistochemistry (Frozen sections) - AEC Substrate Kit (ab64252)Image from Trus et al., Viruses; 12(5):579. doi: 10.3390/v12050579. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - AEC Substrate System (Ready to Use) (ab64252)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - AEC Substrate System (Ready to Use) (ab64252)
    Human colon carcinoma stained with anti-Keratin 20 antibody using AEC Substrate System (Ready to Use) (ab64252).

Protocols

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.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (13)

Publishing research using ab64252? Please let us know so that we can cite the reference in this datasheet.

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
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-4 of 4 Abreviews or Q&A

Question

Thank you for your prompt response. Do you recommend any buffer to go with the heat mediated retrieval, citrate, Tris/EDTA? DAB detection system would suit, I see you have detection kits such as https://www.abcam.com/Mouse-and-Rabbit-Specific-HRPDAB-ABC-detection-IHC-kit-ab64264.html that include secondary antibody.

I will performing immunohistochemisty using DSG3 (3G133), p63 (4A4) and RTNK2 (2C2) and if possible would like a detection kit to suit all. Would https://www.abcam.com/Mouse-and-Rabbit-Specific-HRPDAB-ABC-detection-IHC-kit-ab64264.html be suitable for this?

Read More

Abcam community

Verified customer

Asked on Dec 06 2012

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

Abcam Scientific Support

Answered on Dec 06 2012

Question

I am enquiring about Rhotekin 2 primary antibody for use with IHC-P. I see you have an antibody clone 2C2, can you provide any information on appropriate epitope retrieval methods and detection kits or secondary antibody that would be suitable for use with this antibody?

Read More

Abcam community

Verified customer

Asked on Dec 05 2012

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

Abcam Scientific Support

Answered on Dec 05 2012

Question

Please confirm me whether I can use Mouse and Rabbit Specific HRP/DAB (ABC) detection IHC kit (ab64264) and AEC Substrate System (Ready to Use) (ab64252)
for rat sections. Most of the research is carried out in rat and mice,it will be helpful for the researchers, if you provide the kits that will work in rat, mice, human, rabbit samples.

Read More

Abcam community

Verified customer

Asked on Dec 05 2012

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

Abcam Scientific Support

Answered on Dec 05 2012

Question

Hello,I want to buy substrats for Alkaline phosphatase and HRP for double elispot test.I have seen that you have for alkaline phosphatase  BCIP /NBT :ab7413ab7468for HRP AEC :ab103742ab64252but it is only mentionned for IHC.Can I use these reagents for elispot?Do they need to be filtered or not?What is the protocol (volume to dispense, dilution...?)Thanks for your answer.

Read More

Abcam community

Verified customer

Asked on Sep 08 2011

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.

Read More

Abcam Scientific Support

Answered on Sep 08 2011

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