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An Introduction to Immunohistochemistry

**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 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
  • Troubleshooting
  • Some special challenges
  • Direct staining in IHC

Direct staining in Immunohistochemistry

The application of direct staining in immunohistochemistry (where the primary antibody is directly conjugated to HRP or Alkaline Phosphatase) has a number of advantages including:

  • Significant savings in time and reagents – no secondary antibodies required and fewer wash steps
  • Simple dual staining
  • Simple mouse on mouse staining

However, availability of directly conjugated antibodies for IHC staining is highly restricted. Our Lightning-Link® antibody labeling kits enable the direct conjugation of primary antibodies to HRP, Alkaline Phosphatase and a number of other detection labels, with only 30 seconds hands-on time and with 100% recovery of materials.

The below comparison shows anti-CD20 antibody L26 labelling paraffin-embedded bone marrow, both in a standard indirect technique with Dako Envision as the detection system (used as a control) and the same antibody directly labelled with HRP using Lightning-Link®.

There are examples of the antibody at different concentrations, and with different HRP labelling ratios – all show clear staining of the B lymphocytes, without background staining.

Figure 1. Anti-human CD20 (clone L26) was conjugated to HRP using Lightning-Link® kit, and used in IHC in comparison to a traditional indirect technique.

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