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Establishment, routine, evaluation - Immunohistochemistry for tissue sections or cell cultures

With high-quality equipment and many years of experience, our service develops and validates reproducible routine protocols (SOPs) for immunohistochemistry (IHC) on paraffin, vibratome, cryo and plastic sections as well as on cell cultures or tissue models.

We establish detection reactions with classical colour substrates (e.g. DAB) as well as with modern fluorescent dyes of your choice. We have many years of IHC expertise in human, zoological and botanical tissues with numerous mono- and polyclonal antibodies.

Preparation and fixation of the tissue

  • gentle sampling and preparation
  • Fixation with buffered paraformaldehyde (PFA)
  • Shock freezing in liquid nitrogen
  • Cold embedding in plastics
  • Warm embedding in paraffin below 58°C

Antigen retrieval

Various methods for antigen demasking:

  • Heat treatment in steam cooker and microwave oven
    • heat induced epitope retrival (HERE)
  • Enzymatic unmasking
    • proteolytic induced epitope retrieval (PIER)
      • Proteinase K
      • Trypsin
      • Pepsin
      • Hyaluronidase

Established verifications

  • Cluster of differentiation (CD)
  • Ki-67
  • cytokeratins
  • Collagen and elastin
  • Vasopressin and Oxytocin
  • HMB45 and MelanA
  • Apoptosis Detection (TUNEL)
  • TRAP & AP (Leukocytes)

Further information

Range of methods

All common methods for best results:

  • Direct method
  • Indirect method
    • Use of polymers (Enhanced One Step Method, EPOS)
    • (Strept) avidin biotin methods
    • PAP, APAAP method

Further information

Single markings with chromogens and fluorochromes

Effective and fast immune detection with peroxidase or alkaline phosphatase, with or without reinforcement and counterstaining for orientation in the tissue

Multiple markings with fluorescent and chromogenic dyes

brilliant and consistent results with double and multiple markings using the latest dyes

Establishment studies

Establishment service for you reproducible routine protocols (SOPs) for the IHC on paraffin, cryo and plastic sections as well as on cell cultures.