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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

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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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…

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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

        Download your copy Expedeon 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…

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Guide to labeling your primary antibody

August 23rd, 2017

        Download a free copy! 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…

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Immunohistochemistry

August 23rd, 2017

        Free copy! 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…

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Antibody labeling protocol video

May 7th, 2018

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

**This webinar was produced by Innova Biosciences which is now fully integrated with their company, Expedeon Ltd., and have taken the Expedeon Ltd name and logo. Immunohistochemistry - The basic principles and how to simplify your staining procedures This webinar on immunohistochemistry discusses assay principles, general protocol and tips and…

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

January 20th, 2014

Webinars

**This webinar was produced by Innova Biosciences which is now fully integrated with their company, Expedeon Ltd., and have taken the Expedeon Ltd name and logo.   Dr Nick Gee, the CEO and CSO of Expedeon, explains the benefits of using directly conjugated antibodies and the various approaches and techniques…

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