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    products/reagents/proteinase-k-antigen-retrieval-solution-ab64220.pdf

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Cell Biology Proteolysis / Ubiquitin Proteolytic enzymes Other proteases
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Proteinase K Antigen Retrieval Solution (ab64220)

  • Datasheet
  • SDS
Reviews (1)Q&A (6)References (18)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Proteinase K (ab64220)

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    Overview

    • Product name

      Proteinase K Antigen Retrieval Solution
    • Tested applications

      Suitable for: IHC-P, In situ hybridizationmore details
    • General notes

      ab64220 is a single-component, ready-to-use proteinase K solution for use in protease-induced antigen retrieval during IHC (PIER) and pre-treatment for in situ hybridization.
       

       

      Staining Protocol:

      1. IHC procedure: Incubate tissue section for 5 to 7 minutes at room temperature; Rinse and proceed with immunostaining.
      2. General ISH procedure: Incubate tissue section for 15 minutes at 37oC; Rinse and proceed with hybridization.

      Other products for IHC

      Other enzymes for antigen retrieval include: Pepsin Solution ab64201 and Trypsin Antigen Retrieval Kit ab259256.

      Find more 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 -20°C. Stable for 12 months at -20°C
    • Storage buffer

      pH: 7
      Preservative: 0.1% Sodium azide
      Constituents: Tris buffer, 10% Glycerol (glycerin, glycerine)
    • Concentration information loading...
    • Research areas

      • Cell Biology
      • Proteolysis / Ubiquitin
      • Proteolytic enzymes
      • Other proteases
      • Epigenetics and Nuclear Signaling
      • Assays & Kits
      • ChIP Related Products
      • Kits/ Lysates/ Other
      • Kits
      • IHC tools
      • Antigen retrieval reagent
      • PIER
    • Relevance

      Proteinase K is an endolytic protease that cleaves peptide bonds preferentially next to the carboxyl group of N-substituted hydrophobic aliphatic and aromatic amino acids. Proteinase K is classified as a serine protease and exhibits broad substrate specificity. It can hydrolyze keratin; hence the name. Proteinase K is used for the digestion of proteins on tissue sections as a pre-treatment to aid in the preparation of mRNA and its detection by in situ hybridisation methods using oligonucleotide probes.
    • Alternative names

      • Endopeptidase K
      • Tritirachium alkaline proteinase

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab64220 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.
    In situ hybridization
    Use at an assay dependent dilution.
    Notes
    IHC-P
    Use at an assay dependent dilution.
    In situ hybridization
    Use at an assay dependent dilution.

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Proteinase K (ab64220)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Proteinase K (ab64220)
      ab64220 Proteinase K used with aa anti-Pan CK antibody on Human skin

    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 (18)

    Publishing research using ab64220? Please let us know so that we can cite the reference in this datasheet.

    ab64220 has been referenced in 18 publications.

    • Mold MJ & Exley C Aluminium co-localises with Biondi ring tangles in Parkinson's disease and epilepsy. Sci Rep 12:1465 (2022). PubMed: 35087154
    • Van Den Berge N  et al. Ageing promotes pathological alpha-synuclein propagation and autonomic dysfunction in wild-type rats. Brain 144:1853-1868 (2021). PubMed: 33880502
    • Khoury EE  et al. Distribution of Cardiac and Renal Corin and Proprotein Convertase Subtilisin/Kexin-6 in the Experimental Model of Cardio-Renal Syndrome of Various Severities. Front Physiol 12:673497 (2021). PubMed: 34733169
    • Khoury EE  et al. Pulmonary, cardiac and renal distribution of ACE2, furin, TMPRSS2 and ADAM17 in rats with heart failure: Potential implication for COVID-19 disease. J Cell Mol Med 25:3840-3855 (2021). PubMed: 33660945
    • Kutlu T  et al. Tyrosol retards induction of fibrosis in rats. J Food Biochem 45:e13965 (2021). PubMed: 34636053
    View all Publications for this product

    Customer reviews and Q&As

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

    1-7 of 7 Abreviews or Q&A

    fast and easy

    Excellent Excellent 5/5 (Ease of Use)
    Abreviews
    Abreviews
    Proteinase K digestion has worked well for cytokeratin 20 (ab854) staining.
    The enzyme is ready to use and requires a maximum of 15 minutes for digestion.
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Feb 08 2018

    Question

    Phone call requesting information on how the product needs to be diluted prior to use.

    Read More

    Abcam community

    Verified customer

    Asked on May 20 2013

    Answer



    I can confirm that the Proteinase K (ab64220) is provided at 0.2 mg/ml, this is 200 ug/ml proteinase K. This can be used directly or diluted further if neccessary.

    Read More

    Abcam Scientific Support

    Answered on May 20 2013

    Question

    What is the proteinase K concentration of ab64220?

    Read More

    Abcam community

    Verified customer

    Asked on Sep 19 2012

    Answer

    Thank you for contacting us.

    The concentration of proteinase K is approximately 200 ug/ml. For the 10ug/ml working dilution that you said you want for in situ hybridization, you will need to dilute it 1/20.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Sep 19 2012

    Question

    Hi Kate,

    Thank you for your response.

    I am not quite sure what you mean by ‘With regards to the proteinase K, that should be fine.’

    The customer would like help finding a suitable proteinase K.

    Are you able to help look with the information I have provided you before?

    Thanks again J

    Kind regards,

    Read More

    Abcam community

    Verified customer

    Asked on Apr 05 2012

    Answer

    Thank you for your reply.

    In theory, the proteinase K could also be used in DNA preparation. However, please not that this has not been tested. This reagent is sold as an antigen retrieval buffer.

    I hope this will be helpful. If you have any further questions, please do not hesitate to contact us.

    Read More

    Abcam Scientific Support

    Answered on Apr 05 2012

    Question

    A another required information: How many units "Proteinase K (ab64220) has"? Thank you in advance.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 25 2012

    Answer

    Thank you for contacting Abcam. I have inquired to the lab about the concentration of Proteinase K in this product and have been told it is w/v. This product may be used directly as an antigen retrieval system for IHC. Incubate for 5-7 minutes at room temperature. Rinse and proceed with the IHC protocol. I hope this information is helpful. Please let me know if you have any questions.

    Read More

    Abcam Scientific Support

    Answered on Jan 25 2012

    Question

    Dear Madam / Sir, IHC protocol in https://www.abcam.com/ps/pdf/protocols/ihc_p.pdf link have some absent information. In the document; When it is mention about enzymatic retrieval, it is written "for pepsin and proteinase K, see Section G" But, in the section isn't include these recipes. By the way, we would like to suggest Proteinase K (ab64220) to a customer. But we need its IHC application information: what is recommended Proteinase K concentration for IHC generally?

    Read More

    Abcam community

    Verified customer

    Asked on Jan 25 2012

    Answer

    Thank you for contacting us.

    Thank you for making us aware of the oversight in the IHC handbook. I have been searching through our recommended protocols for antigen retrieval using Proteinase K but have not found an Abcam recommendation for the appropriate concentration. I have found been able to find other sites recommending a 20ug/ml solution and would recommend that you suggest that level to the customer. I have included a link below:

    http://www.ihcworld.com/_protocols/epitope_retrieval/proteinase-k.htm

    I also have sent a request for more information about the concentration/units of Proteinase K in ab64220 and will contact you as soon as I hear back from the lab.


    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Jan 25 2012

    Question

    Dears,   How many tests this can do? Proteinase K (ab64220)   Can it do up to 200 tests? Thanks !   --   Kindest regards,    

    Read More

    Abcam community

    Verified customer

    Asked on Dec 07 2011

    Answer

    Thank you for your enquiry. The number of tests that can be done with the 4 ml of proteinase K provided depends on the customers own procedure.  It is provided ready to use, and will not require diluting. If you are using this for antigen retrieval on slides, then the number of tests you can do will depend on how much you put on each slide. For example, if you put 100 ul on each slide, then you would be able to do 40 tests with 4 mls. I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

    Read More

    Abcam Scientific Support

    Answered on Dec 07 2011

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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