Creative Biolabs is offering the most comprehensive services for antibody development projects. With strict regulation and effective execution, we are dedicated to providing the most valuable solutions to complete your projects.
HighlightsCreative Biolabs is offering the most comprehensive services for antibody development projects. With strict regulation and effective execution, we are dedicated to providing the most valuable solutions to complete your projects.
HighlightsCreative Biolabs is offering the most comprehensive services for antibody development projects. With strict regulation and effective execution, we are dedicated to providing the most valuable solutions to complete your projects.
HighlightsCreative Biolabs is offering the most comprehensive services for antibody development projects. With strict regulation and effective execution, we are dedicated to providing the most valuable solutions to complete your projects.
HighlightsWith over a decade of experience in phage display technology, Creative Biolabs can provide a series of antibody or peptide libraries that are available for licensing or direct screening. These ready-to-use libraries are invaluable resources for isolating target-specific binders for various research, diagnostic or therapeutic applications.
HighlightsCreative Biolabs has established a broad range of platforms for developing novel antibodies or equivalents. These cutting-edge technologies enable our scientists to meet your demands from different aspects and tailor the most appropriate solution that contributes to the success of your projects.
HighlightsWith deep understanding in antibody-related realms and extensive project experience, Creative Biolabs offers a variety of references to help you learn more about our capacities and achievements, including infographic, flyer, case study, peer-reviewed publications, and all kinds of knowledge that can assist your projects. You are also welcome to contact us directly for more specific solutions.
HighlightsGet a real taste of Creative Biolabs, one of the most professional custom service providers in the world. We are committed to providing highly customized comprehensive solutions with the best quality to advance your projects.
HighlightsImmunohistochemistry (IHC) is a laboratory technique used to visualize and detect specific proteins or antigens in tissue samples. It combines principles from immunology and histology, allowing researchers and medical professionals to study the distribution, localization, and expression levels of target proteins within biological tissues. By using antibodies that specifically bind to the protein of interest, IHC provides valuable information about the presence and cellular localization of these proteins, helping to understand various biological processes and disease mechanisms.
The protocol for performing immunohistochemistry can vary slightly depending on the target antigen and the specific antibody being used. However, the general steps involved in the IHC protocol are as follows:
Fig 1. Protocol of immunohistochemistry.
The choice of tissue sample depends on the research question or diagnostic purpose. Tissues can be obtained from surgical specimens, biopsies, or animal models. For formalin-fixed paraffin-embedded (FFPE) tissue, the samples are fixed in formaldehyde (formalin) to preserve the tissue structure and antigens. Frozen sections are obtained by embedding the tissue in a cryoprotective medium (such as OCT compound) and rapidly freezing it using liquid nitrogen or a cryostat.
Paraffin embedding is a common method to preserve tissues, but it hinders antibody penetration. Deparaffinization involves removing the paraffin by incubating slides in xylene or a safer alternative such as xylene substitutes or graded alcohols.
After deparaffinization, rehydration is carried out by passing the slides through a series of decreasing concentrations of alcohol (e.g., 100%, 95%, and 70% ethanol) to restore tissue hydration and improve antibody access.
Formalin fixation can cause cross-linking of proteins, which can mask or alter antigenic epitopes. Antigen retrieval methods, such as heat-induced epitope retrieval (HIER) or enzymatic retrieval, help unmask the antigens, making them accessible to the antibodies.
To prevent nonspecific binding of antibodies, the tissue sections are incubated with a blocking solution. Common blocking agents include 5% bovine serum albumin (BSA) or serum from the same species as the secondary antibody.
The primary antibody is specific to the target antigen of interest. The incubation time and temperature are optimized for each primary antibody and can vary from a few hours to overnight at 4°C or room temperature.
After incubation with the primary antibody, the slides are washed several times with an appropriate buffer (e.g., phosphate-buffered saline, PBS) to remove unbound antibodies.
The secondary antibody is raised against the species of the primary antibody and is conjugated with a label. Common labels include enzymes (e.g., horseradish peroxidase, HRP) for chromogenic detection or fluorescent dyes for fluorescence-based detection. (Creative Biolabs can offer various antibody labeling services to help our customers obtain their very own labeled antibodies.)
Some IHC assays may require an amplification step to enhance the signal. Amplification methods can involve using a secondary antibody against the primary antibody or using polymer-based amplification systems.
Similar to the previous washing step, the slides are washed to remove unbound secondary antibodies or amplification reagents.
For chromogenic detection, a chromogenic substrate specific to the enzyme on the secondary antibody is added. The enzyme catalyzes a reaction that produces a visible color change at the site of antigen-antibody binding. For fluorescence-based detection, counterstaining with a fluorescent dye that binds to DNA (e.g., DAPI) is commonly used to visualize cell nuclei. The slides are then mounted with coverslips using fluorescence mounting media.
The slides are examined using a bright-field or fluorescence microscope. Images are captured and analyzed to determine the localization and intensity of the target antigen within the tissue.
Immunohistochemistry (IHC) is a valuable technique used in various fields of biomedical research, diagnostics, and clinical practice. It involves the use of specific antibodies to detect and visualize specific proteins or antigens in tissues. Here are some common applications of immunohistochemistry:
IHC allows researchers and pathologists to determine the cellular and tissue localization of specific proteins. By staining tissue sections with antibodies targeting particular proteins, they can identify the precise location of these proteins within different cell types and tissue structures. This information is crucial for understanding the functional roles of proteins in normal and diseased tissues.
IHC can provide semi-quantitative or quantitative data on protein expression levels. By analyzing the intensity of staining or the number of positive cells, researchers can compare protein expression between different samples, conditions, or disease stages. This quantitative aspect of IHC is valuable in identifying potential diagnostic or prognostic biomarkers.
Beyond tissue localization, IHC can reveal the subcellular localization of proteins. For example, it can show whether a protein is present in the cell nucleus, cytoplasm, or specific organelles. This knowledge is essential for understanding intracellular signaling pathways and protein trafficking.
Advanced techniques such as dual and multiplex IHC enable the simultaneous visualization of multiple proteins within the same tissue section. By using different-colored labels or fluorescent dyes, researchers can study interactions between proteins, identify co-expression patterns, and gain a more comprehensive view of the molecular context within tissues.
IHC is particularly useful in characterizing tumor heterogeneity, which refers to the diversity of cell types and protein expression patterns within a tumor. Different regions of a tumor may express distinct biomarkers, affecting treatment response and patient outcomes. IHC can help identify and understand these variations.
In cancer research, IHC aids in identifying predictive and prognostic biomarkers that guide treatment decisions. Predictive biomarkers indicate the likelihood of a positive response to a specific therapy, while prognostic biomarkers provide insights into a patient's overall disease outcome.
IHC is crucial in validating potential therapeutic targets identified through other research methods like genomics or proteomics. By confirming the presence of the target protein in relevant tissues, researchers can assess its suitability for drug development.
A cutting-edge technique called Imaging Mass Cytometry combines IHC with mass spectrometry to provide highly detailed spatial information on protein expression within tissues. This technology enables researchers to analyze numerous proteins simultaneously at subcellular resolution.
IHC is employed to identify and study infectious agents, such as viruses or bacteria, in tissues. It helps researchers understand the tissue tropism of pathogens and their effects on host tissues.
IHC serves as a crucial tool for validating the specificity and sensitivity of new antibodies or diagnostic assays. It ensures the accuracy and reliability of these reagents for further research or clinical use.
All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.
