Journal: Drug Invention Today

Article Id: JPRS-Pcol-00004348
Title: Standard operating protocol for immunohistochemical epithelial markers
Category: Pharmacology
Section: Research Article
  • Abstract
  • Audio Abstract
  • Authors
  • Pdf File
  • Citation
  • My Reference
  • Methodology
  • Abstract

    Introduction: Immunohistochemistry (IHC) is an important application of monoclonal and polyclonal antibodies to determine the tissue distribution of an antigen of interest in health and disease. IHC is widely used for diagnosis of cancers; specific tumor antigens are expressed de novo or up-regulated in certain cancers. Cytokeratins (CKs) are intermediate filaments of the cytoskeletons found in all types of epithelial cells. Thus, they are specific for epithelial cell lineage. CKs are over-expressed in Oral squamous cell carcinoma (OSCC) compared to normal mucosa because cancerization of normal oral epithelium could lead to variation of degree of differentiation of epithelium. Epithelial membrane antigen (EMA), a product of Mucin 1 gene, is widely distributed in epithelial and mesothelial tissues and tumors arising from them. Squamous cell carcinomas show positive EMA staining of cytoplasmic membranes. Aim: The aim of the study was to formulate a standard operating protocol for immunohistochemical epithelial markers, CK, and EMA. Materials and Methods: Two tissue blocks of known histopathological diagnosis were procured from the archives of the department of oral and maxillofacial pathology. Four sections of 2.5 µm were cut from each block. Eight lysin coated slides were thus prepared and incubated overnight. The slides used were positive controls of CK and EMA. Three variations were tried and compared with the normal immunohistochemical protocol. Results: The staining intensity, uniformity of staining intensity, and specificity of variation 3 were better, compared to the normal protocol for CK and EMA. Variation 3 showed minimal background staining compared to other variations. Conclusion: It can be concluded that the variation in which deparaffinization was done for 35 min, antigen retrieval for 1 whistle, incubation time increased to 80 min, and 3,3’ Diaminobenzidine chromogen to buffer ratio altered to 1:25, showed better result and can be used as a standard operating protocol for immunohistochemical epithelial markers, CK, and EMA.

  • Abstract Audio

    No Audio file found

  • About the authors and Affiliations

    Author(s) Name:

    Rahul Mohandas*, Pratibha Ramani, Herald J. Sherlin, S. Gheena, Abilasha Ramasubramanian, Gifrina Jayaraj, K. R. Don, Archana Santhanam

    Affiliation(s) Name:

    Department of Oral and Maxillofacial Pathology, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai, Tamil Nadu, India

    *Corresponding author: Rahul Mohandas, Department of Oral and Maxillofacial Pathology, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Velappanchavadi, Chennai - 600 077, Tamil Nadu, India.

  • View Article File in pdf format.

    Article File
  • View Article Citation Here.

    0 View More
  • How to Cite my Article.

    Author:

    Rahul Mohandas*, Pratibha Ramani, Herald J. Sherlin, S. Gheena, Abilasha Ramasubramanian, Gifrina Jayaraj, K. R. Don, Archana Santhanam

    Title:Standard operating protocol for immunohistochemical epithelial markers
    Journal:Drug Invention Today
    Vol(issue):11 (November )
    Year:2019
    Page No: (2745-2750)
  • Experimental Methods Keywords

    Methodology: Immunohistochemistry, Protocol, Standard operating
    Research Materials:Two tissue blocks

Keywords

Cytokeratin Epithelial marker Epithelial membrane antigen Immunohistochemistry Protocol Standard operating

Our Services

Most Downloaded List