Immunohistochemistry-Paraffin protocol for Mesothelin Antibody (NBP2-75255):
Specimen Collection and Preparation
Tissues fixed in 10% formalin are suitable for use prior to paraffin embedding. Consult references (Kiernan, 1981: Sheehan & Hrapchak, 1980) for further details on specimen preparation.
The user is advised to validate the use of the products with their tissue specimens prepared and handled in accordance with their laboratory practices.
Staining Procedure
Refer to the following for conditions specifically recommended for this antibody:
Positive Control - Mesothelioma
Concentrated Dilution - 1:10-1:15
Pretreatment - Buffer pH 8.0
Quality Control
Refer to CLSI Quality Standards for Design and Implementation of Immuno-histochemistry Assays; Approved Guideline-Second edition (I/LA28-A2) CLSI Wayne, PA USA (www.clsi.org). 2011.
Troubleshooting
Contact Novus Technical Support (888.506.6887) to report unusual staining.
Cellular Localization
Cell membrane
Limitations of the Procedure
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. Tissue processing and handling prior to immunostaining can also cause inconsistent results. Variations in fixation and embedding or the inherent nature of the tissue may cause variations in results (Nadji and Morales, 1983).
Endogenous peroxidase activity or pseudoperoxidase activity in erythrocytes and endogenous biotin may cause non-specific staining depending on detection system used.
Tissues containing Hepatitis B surface Antigen (HBsAg) may give a false positive with horseradish peroxidase systems (Omata et al, 1980). Improper counterstaining and mounting may compromise the interpretation of results
