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Validating Antibody Specificities for Immunohistochemistry by Protein Blotting Methods.
Biji T Kurien1,2,3, R Hal Scofield4,5,6
1Arthritis and Clinical Immunology Program, Oklahoma Medical Research Foundation, 825 NE 13th Street, Oklahoma City, OK, 73104, USA. biji-kurien@omrf.ouhsc.edu.
Methods in Molecular Biology (Clifton, N.J.)
|February 11, 2017
Summary
Western blotting (protein blotting) confirms antibody specificity for research and diagnostics. This powerful immunodetection method is crucial for validating antibodies used in immunohistochemistry and detecting low-abundance proteins.
Area of Science:
- Biochemistry
- Molecular Biology
- Immunology
Background:
- Immunoblotting techniques, including western blotting (protein blotting), are vital for validating antibody-based detection methods.
- Enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry (IHC) rely on specific antibodies for accurate results in research and diagnostics.
Purpose of the Study:
- To highlight the critical role of western blotting in confirming antibody specificity for immunohistochemical studies.
- To emphasize western blotting as an essential tool for validating antibodies used in various research and diagnostic applications.
Main Methods:
- Western blotting (protein blotting) involves electrophoretic separation of proteins followed by immunodetection.
- The procedure allows for the simultaneous resolution and detection of multiple immunogenic antigens within a sample using specific antibodies.
Main Results:
- Western blotting provides a powerful method for the immunodetection of proteins, particularly those present in low abundance.
- The technique is instrumental in confirming the specificity of antibodies, a critical step for reliable immunohistochemistry.
Conclusions:
- Western blotting is an indispensable technique for antibody validation in immunohistochemistry.
- The evolution of protein blotting offers researchers diverse approaches to ensure antibody accuracy and reliability in scientific investigations.
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