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Pepsin Solution (Antigen Retrieval) (ab64201)

Submit a review Q&A (4)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Pepsin Solution (ab64201)

    Overview

    • Product name

      Pepsin Solution (Antigen Retrieval)

    • Tested applications

      Suitable for: IHC-Pmore details

    • General notes

      Pepsin for protease-induced antigen retrieval (PIER) during IHC procedures (ab64201). Ready-to-use formulation. Tris buffer pH 2.0

      Staining Protocol:

      1. Incubate tissue section for 10 minutes at 37°C.
      2. Rinse and proceed with immunostaining.

       Pepsin is a commonly used digestive enzyme for immunohistochemical procedures.

      Other products for IHC

      Other enzymes for antigen retrieval include: Proteinase K Antigen Retrieval Solution ab64220 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

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab64201 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 concentration.
    Notes
    IHC-P
    Use at an assay dependent concentration.

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Pepsin Solution (ab64201)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Pepsin Solution (ab64201)
      ab64201 Pepsin Solution used with Laminin on Human Kidney

    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

    References (0)

    Publishing research using ab64201? Please let us know so that we can cite the reference in this datasheet.

    ab64201 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-4 of 4 Abreviews or Q&A

    Question

    Dear Sir/Madam,



    I wonder if you could tell me whether this pepsin solution is human pepsin and whether it would be suitable for trypsin digestion to assess human pepsin peptides by mass spectrometry?



    Thank you for your help,





    Best Wishes

    Read More
    Answer

    Thank you for your inquiry.

    I assume your question is regarding ab64201 (Pepsin Solution)?

    Unfortunately, ab64201 is of porcine origin not of human.

    We also do not have any data about this product that tested any other application than pretreatment of tissue for IHC.

    I am sorry to confirm that we do not have any human pepsin in our catalog.

    I hope this information was nevertheless helpful and wish you good luck in your research.

    Read More

    Question

    IHC-Fr with adult rat brain (hippocampus)
    no signal at all
    20 um sections
    fixation: PFA 4% overnight
    Ab: ab6994: 1/200, 1/500, 1/800 o/n 4 degree
    ab104225: 1/500, 1/1000, 1/2000 o/n 4 degree
    block: none
    secondary: 1/200

    suggestions: try AR (enzymatic), check if secondary works (fluorescent plate reader or fluorescence micrscope, with other primary Ab)

    Read More
    Answer

    Thank you for contacting us.
    I am sorry to hear that the antibodie might not be working as expected.

    As we discussed over the phone the following troubleshooting tips might help in finding the cause of the problem:

    1) Try antigen retrieval (enzymatic) using one of the products we discussed (ab64201, ab64220, ab64205 or ab970).

    2) Check if secondary antibody works (e.g. with fluorescent plate reader or fluorescence micrscope, or with another primary Ab)

    Please let me know if and how your results improving.
    I wish you good luck and look forward to hear back from you.

    Use our products? Submit an Abreview. Earn rewards!
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    Question

    My question is about the pepsin concentration that can be used for nucleic acid retrieval (e.g. in FISH).
    I plan using pepsin for permeabilisation of paraformaldehyde fixed frozen sections for RNA-FISH.
    I have no personal experience in this technique yet.
    Most of available publications recommend pepsin treatment and stress that duration/strength of permeabilisation is important. However, I am struggling to find proper description of pepsin concentration.
    For instance, one of very good papers in nature Methods indicates pepsin concentration as “0.01% pepsin (Sigma) in 0.1 M HCl” (Larsson 2010).
    However, Sigma sells four different pepsin powders with activity ranging from 600 to 4,500 units/mg protein (P7125, P7000, P7012, P6887). I prefer not to dissolve pepsin myself from powder.
    This is why I noted your 20% pepsin solution (cat no ab64201).
    I supose that this is is a concentrated stock solution. Is it correct? If yes, then I consider using this Abcam’s concentrated stock solution.
    However, I am confused about the range of concentrations that I need to try.
    My main questions are:
    1) What working concentration would you recommend for 10-30 min incubation for RNA-FISH permeabilisation on paraformaldehyde fixed frozen sections?
    2) Do you control the activity (in units) of the ab64201 (20% pepsin) ?
    3) Have you checked whether ab64201 is RNAse-free?

    Read More
    Answer



    Unfortunately we do not have any experimental data regarding using the Pepsin Solution ab64201 for RNA-FISH, we do not control the activity of the pepsin and we do not know if it is RNAse-free.

    Please note that the concentration of pepsin in ab64201 is 0.25%, not 20% as currently stated on the product datasheet. This information will be corrected shortly.

    Read More

    Question

    Estimada Ariana,   Buenas  tardes,  podrias  decirme  la concentración que tiene  este  producto,   Pepsin Solution (ab64201)   https://www.abcam.com/Pepsin-Solution-ab64201.html Es para  responder a un  cliente.,  gracias.  

    Read More
    Answer

    Gracias por contactar con Abcam.



    La concentración de este producto es 0,25%.



    Espero que esta información sea útil a usted. Por favor, háganos saber si usted tiene cualquier otra pregunta.

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