Optimization of Immunohistochemical Staining with Anti Protein Gene Product 9,5 (PGP 9,5) Antibodies to Detecting Intraepidermal Nerve Fiber

David Pakaya, Yustina Andwi Ari Sumiwi, Sri Herwiyanti, Rina Susilowati

2020

Abstract

The intraepidermal nerve fibers (INF) are ending branches of the skin sensory nerves. These fibers can be conceived due to the protein gene product 9.5 (PGP 9.5) as a marker recognized by immunohistochemistry. Various studies have visualized these INF with anti-PGP 9.5 antibodies. However, this study differs in immunohistochemical (IHC) staining methods based on the tissue thickness, the antigen retrieval process, and the antibody product used. This study aimed to find an IHC staining optimizer with an anti-PGP 9.5 antibody to detect INF from paraffin blocks. The INF was determined by mice skin biopsy stained with IHC anti-PGP 9.5 antibodies. This procedure was altered in the dilution, duration, incubation temperature of primary antibodies, the tissue's thickness, and the antigen retrieval temperature. We quantitatively analyzed the staining results. Optimization of IHC stain entail of 1:2000 dilution of the primary antibody, the thickness of the tissues were 4 µm, overnight incubation, and low temperature of antigen retrieval. However, the results were inconsistent. The contributing factors that enhance the IHC staining method are thinness of the tissue, low-temperature antigen retrieval, the ratio of antibodies dilution (1: 2000), and incubation overnight at 21°C.

Paper Citation

in Harvard Style

Pakaya D., Ari Sumiwi Y., Herwiyanti S. and Susilowati R. (2020). Optimization of Immunohistochemical Staining with Anti Protein Gene Product 9,5 (PGP 9,5) Antibodies to Detecting Intraepidermal Nerve Fiber.In Proceedings of the 1st Jenderal Soedirman International Medical Conference in conjunction with the 5th Annual Scientific Meeting (Temilnas) Consortium of Biomedical Science Indonesia - Volume 1: JIMC, ISBN 978-989-758-499-2, pages 43-46. DOI: 10.5220/0010487400430046

in Bibtex Style

@conference{jimc20,
author={David Pakaya and Yustina Andwi Ari Sumiwi and Sri Herwiyanti and Rina Susilowati},
title={Optimization of Immunohistochemical Staining with Anti Protein Gene Product 9,5 (PGP 9,5) Antibodies to Detecting Intraepidermal Nerve Fiber},
booktitle={Proceedings of the 1st Jenderal Soedirman International Medical Conference in conjunction with the 5th Annual Scientific Meeting (Temilnas) Consortium of Biomedical Science Indonesia - Volume 1: JIMC,},
year={2020},
pages={43-46},
publisher={SciTePress},
organization={INSTICC},
doi={10.5220/0010487400430046},
isbn={978-989-758-499-2},
}

in EndNote Style

TY - CONF

JO - Proceedings of the 1st Jenderal Soedirman International Medical Conference in conjunction with the 5th Annual Scientific Meeting (Temilnas) Consortium of Biomedical Science Indonesia - Volume 1: JIMC,
TI - Optimization of Immunohistochemical Staining with Anti Protein Gene Product 9,5 (PGP 9,5) Antibodies to Detecting Intraepidermal Nerve Fiber
SN - 978-989-758-499-2
AU - Pakaya D.
AU - Ari Sumiwi Y.
AU - Herwiyanti S.
AU - Susilowati R.
PY - 2020
SP - 43
EP - 46
DO - 10.5220/0010487400430046