Introduction:
In the realm of immunohistochemistry (IHC), achieving accurate and reliable results is paramount for researchers and clinicians alike. Antigen retrieval, a critical step in the IHC process, plays a pivotal role in unmasking antigens and optimizing antibody-antigen interactions. BioGenex’s EZ – Retriever® IR System offers a cutting-edge solution, simplifying antigen retrieval protocols and enhancing the efficiency of this essential procedure. In this article, we delve into the significance of antigen retrieval, explore various antigen retrieval methods, and highlight the benefits of BioGenex’s EZ – Retriever® IR System for IHC applications.
Antigen Retrieval in Immunohistochemistry:
Immunohistochemistry is a method for detecting antigens or haptens in cells of a tissue section by exploiting the principle of antibodies binding specifically to antigens. It is used in many research and clinical laboratories because this technique makes it possible to visualize the distribution and localization of specific cellular components within cells and in proper tissue context. A crucial step in this procedure is ‘Antigen Retrieval’ which involves reversal of cross-linking and leads to unmasking of epitopes. By exposing hidden epitopes, this protocol enables antibodies to bind more effectively, thus enhancing the sensitivity and specificity of IHC staining. A well-optimized antigen retrieval protocol is crucial for obtaining consistent, clean, crisp and reproducible results with immunohistochemical analysis.
Figure: Gastrin on Stomach
Antigen Retrieval Methods – Tailoring to Specific Needs:
Multiple antigen retrieval methods are available, each catering to different tissue types and antigens of interest.
- Heat-induced epitope retrieval (HIER) using heat and pH-adjusted buffers is the most widely employed method.
- Enzymatic retrieval involves the use of enzymes to break down tissue structures and uncover epitopes.
Microwaving, pressure cooking, and water bath methods are also popular options, with variations in temperature and buffer composition allowing researchers to tailor the protocol to their specific IHC requirements.
Antigen Retrieval System + Elegance Solutions – Simplifying the Process:
The high throughput EZ – Retriever® IR System invented by BioGenex is a microwave-based antigen retrieval pre-treatment system using infrared (IR) sensor. This Antigen Retrieval technique breaks the formalin-induced cross-linking bonds between epitopes and unrelated proteins, allowing better penetration of antibody and accessibility of epitopes. It is used for dewaxing, rehydration and antigen retrieval of formalin fixed paraffin embedded (FFPE) tissue sections to enhance exposure of antigenic epitopes. This system resolves the issues of inconsistency due to different types of antigen retrieval methods by applying different microwave power levels, solution volumes and heating time and also facilitates the standardization of dewaxing and antigen protocols for high quality and reproducible staining.
Boiling FFPE tissue sections in buffers such as citrate, EDTA, or Tris, break the methylene bridges formed during fixation and helps recover the masked epitopes. For this purpose, Antigen Retrieval Solutions developed by BioGenex (EZ-AR™ Elegance retrieval solutions) are available with varying pH, Tris and EDTA. They offer one-step dewaxing, retrieval, and rehydration of FFPE tissues for IHC, ISH, FISH & Multiplex staining. These solutions eliminate common issues such as improper heat distribution, buffer evaporation, tissue drying, and boil-overs during antigen retrieval. Elegance solutions are non-hazardous, non-inflammable, odourless, stable up to 1070C and can be reused up to 4 times.
Figure: Antigen retrieval by EZ-AR™ Elegance retrieval solutions from BioGenex
Advantages of EZ – Retriever® IR System for Antigen Retrieval in IHC:
Immunohistochemistry is a powerful technique for visualizing and quantifying protein expression in tissues. However, the presence of masked antigens can lead to false-negative results and compromise the accuracy of IHC analyses. By incorporating BioGenex’s EZ – Retriever® IR System into the IHC workflow, researchers/clinicians can increase staining quality with enhanced sensitivity and specificity, and obtain more reliable data for research and diagnostic purposes.
The EZ – Retriever® IR System is user friendly and high-throughput facilitating standardization of antigen retrieval protocols to produce high-quality and reproducible staining’s on 96 slides in 30 minutes. With exclusive patents issued to BioGenex, this method has been practiced in laboratories throughout the world.
The pre-formulated retrieval EZ-AR™ Elegance retrieval solutions provide consistency and standardization, minimizing the risk of experimental variability. Moreover, the EZ – Retriever® IR System is compatible with a wide range of tissue types and antigens, making it a versatile tool.
Figure: BCL 6 on Tonsil – results with EZ – Retriever® IR System
Conclusion:
Antigen retrieval is an indispensable step in the immunohistochemistry process, allowing researchers and clinicians to obtain accurate and reliable results by unmasking hidden epitopes. BioGenex’s EZ – Retriever® IR System simplifies antigen retrieval protocol, enhancing the efficiency and precision of IHC analyses and interpretation. By incorporating this cutting-edge system into the workflow, can advance the staining quality and enhance the understanding of disease mechanisms and improve patient care in various fields, including pathology, oncology, and biomedical research.