Immunohistochemistry

Principles of Immunohistochemistry (IHC)

Immunohistochemistry (IHC) is the process of using antibodies to detect proteins (antigens) in cells within a tissue section (for instance liver, pancreas or the heart). This tool is used to localize specific antigens in tissue sections with labeled antibodies based on antigen-antibody interactions. The immune reactive products can be visualized by a marker including fluorescent dyes, enzymes in general; radioactive elements or colloidal gold. The IHC principle has been known since the 1930s, but it was not until 1942 that the first IHC study was reported. Scientists used FITC-labeled antibodies to identify Pneumococcal antigens in infected tissue. Since then, improvements have been made in protein conjugation, tissue fixation methods, detection labels and microscopy, making immunohistochemistry an essential tool in diagnostic and research laboratories.

  • Common uses of IHC
  • Fixation
  • Antigen Retrieval
  • IHC Methods
  • Antibody types – monoclonal (mAb) Vs polyclonal (pAb)
  • Antibodies in summary
  • Indirect Vs. Direct Detection Methods
  • Direct Detection Data
  • The Complexity of Immunohistochemistry – Reagent Sourcing and Availability
  • Lightning-Link® – world’s easiest antibody labeling technology
  • IHC reporters
  • Counterstains
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Immunohistochemistry (IHC) – direct labeling

August 28th, 2017

Application notes

Lightning-Link® antibody labeling kits can be used to directly conjugate antibodies for IHC to HRP or other labels. The IHC data below shows that it is possible to obtain very good staining whilst avoiding the use of secondary antibodies, therefore saving significantly in time and reagent costs. Anti-human CD20 (clone L26) was conjugated to HRP…

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Tech tip – Interfering buffer additives

August 23rd, 2017

Lightning-Link® from Innova Biosciences is an innovative technology that enables direct labeling of antibodies, proteins, peptides or any other biomolecule which has free amine groups. These easy to use kits require just 30 seconds hands-on time and, with no separation steps involved, 100% of the biomolecule is recovered. Lightning-Link® conjugates are highly stable, and with…

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A-Z Guide to Fluorochromes

August 23rd, 2017

Download a Free Copy     Lightning-Link® fluorescent antibody and protein labeling kits allow you to covalently label any antibody in under 20 minutes with only 30 seconds hands-on time. We currently have over 35 label in our range of fluorescent dyes. This guide provides a general overview of each fluorochrome in our range including…

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The Biotin Streptavidin Interaction

August 23rd, 2017

The affinity of streptavidin for biotin is the strongest non-covalent biological interaction known, with a dissociation constant (Kd) in the femtomolar range. Each streptavidin monomer can bind one biotin molecule, allowing a streptavidin protein to maximally bind four biotins. The streptavidin-biotin interaction In addition to the strength of the interaction, the binding of streptavidin to…

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Absorption-Emission Table

August 23rd, 2017

Lightning-Link® is an innovative technology that enables direct labeling of antibodies, proteins, peptides or other biomolecules with only 30 seconds hands-on time.

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Simplifying Antibody Conjugation Process

August 23rd, 2017

Antibodies are widely employed in the quantification of antigens in complex biological samples. Using techniques such as Western blotting, ELISA, and immunohistochemistry researchers are able to measure a single antigen, or perhaps a limited number of antigens, in each sample. In the post-genomics era, advances in multiplex immunoassay technologies now allow scores or even hundreds…

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Antibody Purification Guide

August 23rd, 2017

Innova Biosciences specializes in easy to use bioconjugation kits which enable the direct labeling of antibodies or proteins with enzymes, fluorescent labels, biotin, streptavidin, gold nanoparticles, latex beads or oligonucleotides. Unfortunately, many antibodies are provided in buffers which contain additives that are incompatible with labeling technologies, making puri cation a key consideration prior to carrying…

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Guide to labeling your primary antibody

August 23rd, 2017

Antibodies are used to detect and quantify antigens in techniques such as flow cytometry, ELISA, western blotting, immunohistochemistry and lateral flow. The antibody that binds to the antigen is called a ‘primary antibody’ and it confers specificity to the assay. Most types of immunoassays also incorporate a ‘label’ whose purpose is to provide measurability. Assays…

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Immunohistochemistry

August 23rd, 2017

Immunohistochemistry (IHC), or immunohistochemical staining, is a technique which employs antibodies to detect antigens in cells within a tissue section. This application is used to locate specific antigens in tissue sections with labeled antibodies based on antigen-antibody interactions. The immune reactive products can be visualized by a marker including fluorescent dyes, enzymes, radioactive elements or…

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Antibody labeling protocol video

August 7th, 2017

Applications, Videos

Stop using secondary antibodies in immunoassays! Save time and materials by labeling your primary antibodies in under 20 min. For use in ELISA, western blotting, IHC, flow cytometry and many other applications. Scientists love our Lightning-Link® kits because of the remarkable simplicity of the technology. Now they can open a…

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An Introduction to Immunohistochemistry

May 19th, 2014

Webinars

Immunohistochemistry - The basic principles and how to simplify your staining procedures This webinar on immunohistochemistry discusses assay principles, general protocol and tips and hints for simplifying your staining procedures. Other discussion points include: Tissue preparation and fixation and pre-treatment Principles of immunostaining and staining protocols Choosing and using antibodies…

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Methods of antibody conjugation – which one is best for you?

January 20th, 2014

Webinars

Dr Nick Gee, the CEO and CSO of Innova Biosciences, explains the benefits of using directly conjugated antibodies and the various approaches and techniques that researchers can use in their own labs. The webinar covers: Why conjugate your antibody? What are the main chemical approaches? Is there a 'best' method?…

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