Immunohistochemistry (IHC) reagents
Get clear, reliable results with our immunohistochemistry and imaging reagents. These tools help you visualize protein location in tissue sections, giving you insights into distribution and abundance, key to understanding biological processes and disease. It is essential to run controls in IHC staining experiments to confirm that the observed staining pattern is accurate and reliable.
Every product is rigorously tested for sensitivity and specificity, reducing background and improving signal detection. Plus, we provide detailed protocols and expert support so you can achieve optimal staining and accurate interpretation.
Our goal is to help you push the boundaries of research with dependable reagents and guidance.
Antibody diluents
Maintain antibody integrity and ensure consistent staining. Explore our high-quality antibody diluents now.
Antigen retrieval buffers
Optimize the unmasking of antigen sites for improved binding and visualization of target proteins in fixed tissue samples.
IHC detection kits
Comprehensive kits that include all necessary components for robust and reliable immunohistochemical staining in various tissues.
Immunohistochemistry (IHC) and imaging reagents are essential tools in the field of histology and cellular biology, providing detailed visualization of the localization of specific proteins within tissue sections. Using these reagents, researchers can gain invaluable insights into the tissue distribution and abundance of proteins, aiding in the understanding of various biological processes and disease states. Our range of IHC and imaging reagents is designed to enhance the clarity and quality of staining results, which is crucial for accurate data interpretation.
Our offerings include high-quality antibody diluents that maintain the integrity and specificity of antibodies, antigen retrieval buffers that improve antigen visibility in formalin-fixed tissues, and comprehensive IHC detection kits tailored for multiple staining protocols. These products undergo rigorous quality testing to ensure high sensitivity and specificity, thereby minimizing non-specific binding and enhancing signal detection. Moreover, we provide robust technical support and detailed protocol guidance, empowering researchers to achieve optimal results in their IHC experiments. Through these advanced histology reagents and dedicated support, Abcam enables scientists to push the boundaries of their research forward.
Antigen retrieval
Antigen retrieval is a critical step in immunohistochemistry (IHC) that restores antigenicity in formalin-fixed tissues, including fixed paraffin-embedded tissue, which is commonly used for morphological preservation. Methods such as heat induced epitope retrieval (HIER) and enzymatic retrieval address different tissue types and antigens. These techniques break protein cross-links formed during fixation, allowing antibodies to bind effectively.
Read more about protocols for IHC with samples in paraffin.
Blocking and permeabilization
Blocking is an essential step in your IHC experiments as it reduces background signals and false positives. It occurs after sample preparation, but usually before incubation with the primary antibody. As IHC uses antibodies to recognize specific antigens, blocking prevents non-specific binding.
Permeabilization is required for the antibody to access the inside of cells to detect the target antigen, and solvents or detergents are typically used. Solvents are generally recommended for cytoskeletal, viral, and some enzyme antigens. Detergent permeabilization can significantly improve antibody access to antigens in the cytoplasm, on the cytoplasmic face of the plasma membrane, and soluble nuclear antigens.
Detection and chromogens
In IHC, antigens can be detected by either chromogenic or fluorescent methods.
Chromogenic detection relies on enzymes that convert soluble substrates into insoluble chromogenic products. The choice of chromogen is determined by the enzyme used.
Fluorescent detection uses fluorochrome labels, which emit light of a longer wavelength when excited by light of a specific wavelength. It is widely used to visualize multiple antigens simultaneously. The fluorochrome may be conjugated directly to the primary or secondary antibody or streptavidin.
Counterstaining and mounting
Histology, fluorescent, and special stains are used to stain specific cell and tissue structures and microorganisms to help identify the location of primary antibody staining.
- Chromogenic counterstains are used when the primary antibody is visualized using HRP or alkaline phosphatase combined with DAB, AEC, or similar enzyme substrates.
- Fluorescent counterstains are used when the primary antibody staining is visualized with a fluorescent dye conjugated directly to the primary or secondary antibody.
- Special stains are used to stain cell types, microorganisms, and specific proteins, carbohydrates, and metabolites found in the tissue matrix and within cells.
After staining, a mounting medium is used to adhere a coverslip to a tissue section or cell smear. There are two categories of mounting media:
- Organic (or hydrophobic), which can only be used for enzymatic labels where the precipitate formed between the enzyme and the chromogen is not soluble in the organic solvents used during mounting of the tissue.
- Aqueous (or hydrophilic), which is generally suitable for all enzymatic label/chromogen combinations and fluorescent labels.
IHC troubleshooting
Need to get your experiment back on track? Our troubleshooting tips cover solutions to common issues in IHC, including high background, no staining, not getting the desired staining pattern, and more. You can also read our complete IHC guide or contact us for personalized assistance via Chat, email or phone.
Tissue clearing reagents
Explore tissue clearing reagents that make specimens transparent for deep imaging.
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