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TIPS for IHC Antigen Retrieval Optimization Tips

Posted by Immuno Staining on August 6th, 2019

Immunohistochemistry (IHC) is a special test used for the detection of specific antigens (the polypeptide, proteins) on tissue sections. In the biological field, it is quite a powerful tool for pathologists, which utilizes monoclonal and polyclonal antibodies to determine the tissue distribution of an antigen in health and disease samples. IHC has an important role in pathology and it also has plenty of applications in medicine, especially in cancer diagnosis, such as the diagnosis of lymphomas.

Additionally, HC can be particularly helpful in diagnosing abnormalities in diseases such as cancer where tumor samples are available and contextual data is required.

IHC technique

It consists of four main steps:

(1) Tissue fixation- to keep everything in its place

(2) Antigen retrieval- to extend the availability of proteins for detection

(3) Blocking- to minimize pesky background signals

(4) Antibody labelling and visualization- getting pretty pictures.

What is antigen retrieval?

Antigen retrieval is a technique required in most formalin-fixed tissues before immunohistochemical staining. Antigen retrieval (or antigen recovery) is performed to expose or retrieve antigen epitopes which have become masked by the tissue fixation process without modifying the fine structure of the tissue.

Avoiding this step may lead to weak or false negative staining? Antigen retrieval is usually carried out before staining to unmask hidden or denatured target epitopes. There are two methods of antigen retrieval:

  • Heat-induced epitope retrieval (HIER)
  • Proteolytic-induced epitope retrieval (PIER)

And which method to use depends on the tissue type and the primary antibody. Choosing the right IHC antigen retrieval optimization tips is extremely important to get the best outcomes for your program.

Tips for Successful Immunohistochemistry

  • Choose the right antibody and Test it
  • Preparation of Tissue
  • Control
  • Tissue Fixation
  • Antigen Retrieval
  • Blocking
  • Non-specific Binding
  • Detection System Choice
  • Staining

Boster Bio is one of the leading antibody manufacturers specializing in high-sensitivity, high-specificity, ELISA kits, and WB/IHC compatible antibodies. Our antibodies are thoroughly validated for varied applications including IHC, WB, ELISA, and Flow Cytometry.

Boster bio was founded in the year 1993 by famed histologist Steven Xia. We have been here from the last two and half decades for perfecting our techniques and technology and our products have been well-cited in over 23,000 publications.

From the above discussion, we can conclude that in general, the antigen retrieval process is not necessary for frozen sections. However, sometimes acetone- and NBF-fixed frozen sections are required to prevent washout of the target antigen, particularly for soluble antigens. In that case, antigen retrieval may serve for better IHC signals.

To get more understanding about IHC Antigen Retrieval Methods open the official website of Boster Antibody & ELISA Experts. If you ever need support with your experiments, contact the Boster support team any time .for more details please visit our website HERE; https://immunostaining.info/

Also See: Antigen Retrieval, Tissue Fixation, Ihc Antigen, Retrieval Optimization, Retrieval, Ihc, Antigen

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