Heat-induced antigen retrieval restores electrostatic forces: prolonging the antibody incubation as an alternative.

Author(s): Boenisch T

Publication: Appl Immunohistochem Mol Morphol, 2002, Vol. 10, Page 363-7

PubMed ID: 12607606 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of PAI and duration of HIAR on IHC staining on tonsil, prostate, and breast FFPE specimens using a panel of 26 antibodies. HIAR was performed in DAKO Target Retrieval Solution, High pH at 97 degrees C.

   Summary of Findings:

   The author states that, in general, IHC staining for the panel of 26 antibodies was similar after HIAR for 20, 40, or 60 min, and PAI of 60 min, instead of the standard 10 min, resulted in optimal staining intensities even without subsequent HIAR. The antibody dilutions used for PAI generally exceeded the vendor recommended dilutions used after HIAR. However, there were antigen specific exceptions. For CD21 and estrogen receptor (ER), PAI without HIAR resulted in moderate staining intensities, and for Ki-67, no length of PAI resulted in any immunostaining (HIAR was absolutely required). With the exception of progesterone receptor (PR), extending PAI to 120 min instead of 60 min did not further improve immunostaining intensities.

   Biospecimens

   • Tissue - Tonsil
   • Tissue - Breast
   • Tissue - Prostate

   Preservative Types

   • Formalin

   Diagnoses:

   • Normal

   Platform:

   Analyte	Technology Platform
   Protein	Immunohistochemistry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Analyte Extraction and Purification	Antigen retrieval	20 min 40 min 60 min
   Immunohistochemistry Specific	Incubation time	10 min 60 min 120 min

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