A Melanin bleaching method to prevent non-specific immunostaining of chicken feathers

Shahid Nazir; Richard P.G. Charlesworth; Stephen W. Walkden-Brown; Priscilla F. Gerber

doi:10.1016/j.mex.2020.100957

A Melanin bleaching method to prevent non-specific immunostaining of chicken feathers

Shahid Nazir^*
, Richard P.G. Charlesworth
, Stephen W. Walkden-Brown
, Priscilla F. Gerber

^*Corresponding author for this work

Research output: Journal Publications and Reviews › RGC 21 - Publication in refereed journal › peer-review

7 Citations (Scopus)

33 Downloads (CityUHK Scholars)

Abstract

Melanin in pigmented organs like the skin is known to react with 3,3′-diaminobenzidine (DAB) to give a brown colour indistinguishable from the colour that DAB imparts to target antibodies bound to specific antigens. This can lead to false positives in chicken feathers during immunoperoxidase staining. Here, we present a simple, fast and practical method for bleaching chicken feathers which can be applied prior to immunohistochemistry staining without affecting specific antigen-antibody binding. To our knowledge, this is the first report of a melanin-bleaching technique prior to immunoperoxidase staining techniques of chicken feathers for detection of pathogens. Optimisations of the method include:

• Removal of melanin from tissue sections using a short incubation with potassium permanganate followed by incubation with oxalic acid prior to immunostaining for improved specificity.

© 2020 The Authors. Published by Elsevier B.V.
• This technique did not affect the antigenicity of infectious laryngotracheitis virus antigen and did not cause damage or detachment of tissues from the slides.

Original language	English
Article number	100957
Journal	MethodsX
Volume	7
Online published	11 Jun 2020
DOIs	https://doi.org/10.1016/j.mex.2020.100957
Publication status	Published - 2020
Externally published	Yes

Funding

This study was supported in part by Rural Industry Research and Development Corporation (now AgriFutures Australia) grant number PRJ-010639, and by the University of New England postgraduate student research funds.

Research Keywords

Chicken
Diamminobenzidine
Feather follicles
Melanin
Melanin bleaching of chicken feathers
Oxalic acid
Potassium permanganate

Publisher's Copyright Statement

This full text is made available under CC-BY-NC-ND 4.0. https://creativecommons.org/licenses/by-nc-nd/4.0/

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title = "A Melanin bleaching method to prevent non-specific immunostaining of chicken feathers",

abstract = "Melanin in pigmented organs like the skin is known to react with 3,3′-diaminobenzidine (DAB) to give a brown colour indistinguishable from the colour that DAB imparts to target antibodies bound to specific antigens. This can lead to false positives in chicken feathers during immunoperoxidase staining. Here, we present a simple, fast and practical method for bleaching chicken feathers which can be applied prior to immunohistochemistry staining without affecting specific antigen-antibody binding. To our knowledge, this is the first report of a melanin-bleaching technique prior to immunoperoxidase staining techniques of chicken feathers for detection of pathogens. Optimisations of the method include: • Removal of melanin from tissue sections using a short incubation with potassium permanganate followed by incubation with oxalic acid prior to immunostaining for improved specificity. {\textcopyright} 2020 The Authors. Published by Elsevier B.V. • This technique did not affect the antigenicity of infectious laryngotracheitis virus antigen and did not cause damage or detachment of tissues from the slides.",

keywords = "Chicken, Diamminobenzidine, Feather follicles, Melanin, Melanin bleaching of chicken feathers, Oxalic acid, Potassium permanganate",

author = "Shahid Nazir and Charlesworth, \{Richard P.G.\} and Walkden-Brown, \{Stephen W.\} and Gerber, \{Priscilla F.\}",

year = "2020",

doi = "10.1016/j.mex.2020.100957",

language = "English",

volume = "7",

journal = "MethodsX",

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TY - JOUR

T1 - A Melanin bleaching method to prevent non-specific immunostaining of chicken feathers

AU - Nazir, Shahid

AU - Charlesworth, Richard P.G.

AU - Walkden-Brown, Stephen W.

AU - Gerber, Priscilla F.

PY - 2020

Y1 - 2020

N2 - Melanin in pigmented organs like the skin is known to react with 3,3′-diaminobenzidine (DAB) to give a brown colour indistinguishable from the colour that DAB imparts to target antibodies bound to specific antigens. This can lead to false positives in chicken feathers during immunoperoxidase staining. Here, we present a simple, fast and practical method for bleaching chicken feathers which can be applied prior to immunohistochemistry staining without affecting specific antigen-antibody binding. To our knowledge, this is the first report of a melanin-bleaching technique prior to immunoperoxidase staining techniques of chicken feathers for detection of pathogens. Optimisations of the method include: • Removal of melanin from tissue sections using a short incubation with potassium permanganate followed by incubation with oxalic acid prior to immunostaining for improved specificity. © 2020 The Authors. Published by Elsevier B.V. • This technique did not affect the antigenicity of infectious laryngotracheitis virus antigen and did not cause damage or detachment of tissues from the slides.

AB - Melanin in pigmented organs like the skin is known to react with 3,3′-diaminobenzidine (DAB) to give a brown colour indistinguishable from the colour that DAB imparts to target antibodies bound to specific antigens. This can lead to false positives in chicken feathers during immunoperoxidase staining. Here, we present a simple, fast and practical method for bleaching chicken feathers which can be applied prior to immunohistochemistry staining without affecting specific antigen-antibody binding. To our knowledge, this is the first report of a melanin-bleaching technique prior to immunoperoxidase staining techniques of chicken feathers for detection of pathogens. Optimisations of the method include: • Removal of melanin from tissue sections using a short incubation with potassium permanganate followed by incubation with oxalic acid prior to immunostaining for improved specificity. © 2020 The Authors. Published by Elsevier B.V. • This technique did not affect the antigenicity of infectious laryngotracheitis virus antigen and did not cause damage or detachment of tissues from the slides.

KW - Chicken

KW - Diamminobenzidine

KW - Feather follicles

KW - Melanin

KW - Melanin bleaching of chicken feathers

KW - Oxalic acid

KW - Potassium permanganate

UR - https://www.scopus.com/pages/publications/85086661448

UR - https://www.scopus.com/record/pubmetrics.uri?eid=2-s2.0-85086661448&origin=recordpage

U2 - 10.1016/j.mex.2020.100957

DO - 10.1016/j.mex.2020.100957

M3 - RGC 21 - Publication in refereed journal

C2 - 32612940

SN - 2215-0161

VL - 7

JO - MethodsX

JF - MethodsX

M1 - 100957

ER -

A Melanin bleaching method to prevent non-specific immunostaining of chicken feathers

Abstract

Funding

Research Keywords

Publisher's Copyright Statement

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