Overview

  • Product name

    StayRed/AP 30ml (Alcohol & Xylene Compatible)
  • Assay type

    Direct
  • Product overview

    StayRed/AP is a substrate chromogen system designed to be used for either IHC or ISH when using alkaline phosphate detection. StayRed/AP produces a brilliant dark red color. It is also insoluble in organic solvents so sections can be dehydrated in alcohol, cleared in xylene (or a xylene substitute) and permenently mounted.

  • Notes

    Intended Use:

    Substrate/chromogen in conjunction with alkaline phosphatase-based immunostaining or in situ hybridization systems.

     

    Working Solution:

    Aliquot 3mL of StayRed/AP Substrate Buffer in a mixing bottle. Add one drop (~20μL) of StayRed/AP Chromogen. Replace tip, mix, and allow solution to reach room temperature before using.

    Note: The working chromogen-substrate solution should be prepared fresh and used within 20-30 minutes of preparation. Any solution not used during this period should be discarded.

     

    Procedure:

    • After SA-alkaline phosphatase incubation, wash tissue sections with wash buffer.
    • Wipe slides, removing excess buffer. Add enough drops of StayRed/AP working solution to cover tissue sections.
    • Incubate for 5-15 minutes at room temperature. For optimal results, observe reaction under microscope for signal development. Once desired signal to noise ratio is achieved, stop reaction by rinsing slides in wash buffer.
    • Counterstain sections in hematoxylin.
    • Dehydrate sections in alcohol, clear in xylene (or xylene-substitute), and mount in permanent mounting medium.

Properties

Images

  • Formalin fixed paraffin embedded Human tonsil stained with HMW Cytokeratin antibody (ab850) labeled with PermaRed/AP Chromogen (ab103741) produces a brilliant red color.

  • StayRed staining by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).

References

This product has been referenced in:

  • Aznar MA  et al. Immunotherapeutic effects of intratumoral nanoplexed poly I:C. J Immunother Cancer 7:116 (2019). Read more (PubMed: 31046839) »
  • Shami A  et al. Fibromodulin deficiency reduces low-density lipoprotein accumulation in atherosclerotic plaques in apolipoprotein E-null mice. Arterioscler Thromb Vasc Biol 33:354-61 (2013). Read more (PubMed: 23202368) »
See all 2 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer

Thank you for your inquiry.

I can suggest to use a strepavidin that is coupled to an enzyme, for example: ab128985 (Streptavidin Peroxidase (Ready-To-Use)) and combine this with a suitable chromogen, for example: ab94665 (DAB substrate kit)

https://www.abcam.com/index.html?datasheet=128985 (or use the following: https://www.abcam.com/index.html?datasheet=128985).

https://www.abcam.com/index.html?datasheet=94665 (or use the following: https://www.abcam.com/index.html?datasheet=94665).

Another possible combination would be a strepavidin-AP ab128984 (Streptavidin Alkaline Phosphatase (Ready-To-Use)) with a more colourful chromogen like ab103743 (StayBlue/AP 30ml), ab156428 (StayGreen/AP) or ab103741 (StayRed/AP 30ml).

https://www.abcam.com/index.html?datasheet=128984 (or use the following: https://www.abcam.com/index.html?datasheet=128984).

https://www.abcam.com/index.html?datasheet=103743 (or use the following: https://www.abcam.com/index.html?datasheet=103743).

https://www.abcam.com/index.html?datasheet=156428 (or use the following: https://www.abcam.com/index.html?datasheet=156428).

https://www.abcam.com/index.html?datasheet=103741 (or use the following: https://www.abcam.com/index.html?datasheet=103741).

I hope this information is helpful and wish you good luck with your research.

Read More

Answer


The source of StayRed and StayBlue recommend to do sequential staining. Their recommendations on chromogens for double staining are DAB and StayRed (https://www.abcam.com/ab103741). In order to use these chromogens first use DAB, then RED, followed by counter stain ex: Hematoxylin.

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Answer

Thank you for your enquiry. I can confirm that these products can be used together for double staining. However, I can recommend to consider the following: 1. We would suggest to ensure the primary antibodies used in the stains undergo the same conditions for antigen/epitope retrieval. 2. Since the StayRed is a permanent Chromogen (i.e. both alcohol and xylene insoluble) it must be used as the first Chromogen in the double staining protocol. 3. The StayGreen is also alcohol insoluble, however since it is soluble in xylene it must be used with a xylene substitute. The tissue can then be mounted permanently. I was hoping to obtain any images of double staining for you, but regrettably our source does not have these available. I hope this will be helpful to you. Should you have any further questions, please do not hesitate to contact us.

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