Application of the critical molar concentration concept to heat-mediated antigen retrieval in immunohistochemistry.
Previously we have applied Scott's critical molar concentration concept to show that divalent cations, especially Mg2+ may be used to measure the affinity of a known monoclonal antibody for its antigen. In this paper we report the application of this same procedure to a study of a series of antigens (three globular proteins and three intermediate filaments). The concept was applied to samples without any previous treatment or after the application of heat-mediated antigen retrieval (using a pressure cooker). Our findings suggest that heat-mediated antigen retrieval sets free protein side-chain(s) that have been masked by formaldehyde fixation. This is reflected in a higher affinity of the antibody for the antigen in question.
Montero
C
C
Department of Pathology, Hospital Central Dr. Ignacio Morones Prieto, Facultad de Medicina, Universidad Autónoma de San Luis Potosí, Mexico.
Chi-Ahumada
E
E
Chavez-Porras
S
S
Gutierrez
M
M
D000911:Antibodies, Monoclonal; D000915:Antibody Affinity; D002272:Carcinoembryonic Antigen / Q000032:analysis; D000939:Epitopes / Q000032:analysis*; D006358:Hot Temperature; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods*; D007381:Intermediate Filament Proteins / Q000032:analysis; D015636:Magnesium Chloride / Q000737:chemistry*; D017430:Prostate-Specific Antigen / Q000032:analysis; D009418:S100 Proteins / Q000032:analysis
D000911:Antibodies, Monoclonal; D002272:Carcinoembryonic Antigen; D000939:Epitopes; D007381:Intermediate Filament Proteins; D009418:S100 Proteins; D015636:Magnesium Chloride; D017430:Prostate-Specific Antigen