Immunolabeling
Overview[edit | edit source]
Immunolabeling is a technique used in molecular biology and biochemistry to detect specific proteins or antigens in a sample using antibodies. This method is widely used in research and diagnostics to study the presence and distribution of proteins in cells and tissues.
Principles of Immunolabeling[edit | edit source]
Immunolabeling relies on the specific binding of an antibody to its corresponding antigen. The antibody is typically conjugated to a detectable marker, such as a fluorescent dye or an enzyme, which allows for visualization of the antigen-antibody complex.
Types of Immunolabeling[edit | edit source]
There are several types of immunolabeling techniques, including:
- Immunohistochemistry (IHC): Used to detect antigens in tissue sections.
- Immunocytochemistry (ICC): Used for detecting antigens in cell cultures.
- Western blotting: Used to detect proteins in a gel.
Procedure[edit | edit source]
The immunolabeling process generally involves the following steps:
- Sample Preparation: The sample, such as a tissue section or cell culture, is prepared and fixed to preserve the structure and antigenicity.
- Blocking: Non-specific binding sites are blocked to prevent background staining.
- Primary Antibody Incubation: The sample is incubated with a primary antibody specific to the target antigen.
- Secondary Antibody Incubation: A secondary antibody, conjugated to a detectable marker, is applied. This antibody binds to the primary antibody.
- Detection: The marker on the secondary antibody is visualized using appropriate methods, such as fluorescence microscopy or colorimetric detection.
Applications[edit | edit source]
Immunolabeling is used in various fields, including:
- Pathology: To diagnose diseases by detecting abnormal protein expression.
- Neuroscience: To study the distribution of neurotransmitters and receptors.
- Cancer research: To identify tumor markers and study cancer progression.
Advantages and Limitations[edit | edit source]
Immunolabeling offers high specificity and sensitivity, allowing for the detection of low-abundance proteins. However, it requires well-characterized antibodies and can be limited by cross-reactivity and non-specific binding.
Related pages[edit | edit source]
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Contributors: Prab R. Tumpati, MD
