Immunocytochemical methods for demonstrating canine distemper virus antigen in aldehyde-fixed paraffin-embedded tissue

Michael K. Axthelm; Steven Krakowka

doi:10.1016/0166-0934(86)90016-9

Immunocytochemical methods for demonstrating canine distemper virus antigen in aldehyde-fixed paraffin-embedded tissue

Michael K. Axthelm
, Steven Krakowka

Research output: Contribution to journal › Article › peer-review

22 Scopus citations

Abstract

The effects of enzymatic digestion, sodium borohydride reduction, acids used in décalcification procedures and techniques for inactivation of endogenous peroxidase were sequentially evaluated for their effect on the immunoreactivity of canine distemper virus in aldehyde-fixed paraffin-embedded tissue. Enzyme digestion improved immunoreactivity while sodium borohydride reduced background staining. Paraformaldehyde-glutaraldehyde-fixed tissues required thioglycolic acid treatment prior to enzyme digestion and sodium borohydride reduction to obtain results comparable to results obtained in formalin-fixed tissues. Detailed protocols for indirect immunofluorescence and the avidin-biotin-peroxidase complex procedure are provided.

Original language	English (US)
Pages (from-to)	215-229
Number of pages	15
Journal	Journal of Virological Methods
Volume	13
Issue number	3
DOIs	https://doi.org/10.1016/0166-0934(86)90016-9
State	Published - Jun 1986
Externally published	Yes

Funding

The authors thank Nancy Austin, Judy Dubena and Alice Kelley for technical assistance and Virginia Stump for preparation of the manuscript. This work was supported by The State of Ohio Canine Research Fund, The National Institute of Neurological and Communicative Disorders and Stroke, National Institutes of Health Grant SROl-NS14821-15 and U.S. Public Health Service Grant 2 T32 NS07091-06Al 0021.

Funders	Funder number
State of Ohio Canine Research Fund
Author National Institutes of Health National Institutes of Health National Institutes of Health National Institutes of Health The Bev Hartig Huntington's Disease Foundation National Institutes of Health	SROl-NS14821-15
National Institute of Neurological Disorders and Stroke	R01NS014821
U.S. Public Health Service	2 T32 NS07091-06Al 0021

Keywords

aldehyde fixation
canine distemper virus
immunocytochemistry
immunofluorescence technique
immunoperoxidase
technique

ASJC Scopus subject areas

Virology

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10.1016/0166-0934(86)90016-9

Cite this

@article{776faff7f6af452fa6629c1a3491df57,

title = "Immunocytochemical methods for demonstrating canine distemper virus antigen in aldehyde-fixed paraffin-embedded tissue",

abstract = "The effects of enzymatic digestion, sodium borohydride reduction, acids used in d{\'e}calcification procedures and techniques for inactivation of endogenous peroxidase were sequentially evaluated for their effect on the immunoreactivity of canine distemper virus in aldehyde-fixed paraffin-embedded tissue. Enzyme digestion improved immunoreactivity while sodium borohydride reduced background staining. Paraformaldehyde-glutaraldehyde-fixed tissues required thioglycolic acid treatment prior to enzyme digestion and sodium borohydride reduction to obtain results comparable to results obtained in formalin-fixed tissues. Detailed protocols for indirect immunofluorescence and the avidin-biotin-peroxidase complex procedure are provided.",

keywords = "aldehyde fixation, canine distemper virus, immunocytochemistry, immunofluorescence technique, immunoperoxidase, technique",

author = "Axthelm, \{Michael K.\} and Steven Krakowka",

note = "Funding Information: The authors thank Nancy Austin, Judy Dubena and Alice Kelley for technical assistance and Virginia Stump for preparation of the manuscript. This work was supported by The State of Ohio Canine Research Fund, The National Institute of Neurological and Communicative Disorders and Stroke, National Institutes of Health Grant SROl-NS14821-15 and U.S. Public Health Service Grant 2 T32 NS07091-06Al 0021.",

year = "1986",

month = jun,

doi = "10.1016/0166-0934(86)90016-9",

language = "English (US)",

volume = "13",

pages = "215--229",

journal = "Journal of Virological Methods",

issn = "0166-0934",

publisher = "Elsevier B.V.",

number = "3",

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TY - JOUR

T1 - Immunocytochemical methods for demonstrating canine distemper virus antigen in aldehyde-fixed paraffin-embedded tissue

AU - Axthelm, Michael K.

AU - Krakowka, Steven

N1 - Funding Information: The authors thank Nancy Austin, Judy Dubena and Alice Kelley for technical assistance and Virginia Stump for preparation of the manuscript. This work was supported by The State of Ohio Canine Research Fund, The National Institute of Neurological and Communicative Disorders and Stroke, National Institutes of Health Grant SROl-NS14821-15 and U.S. Public Health Service Grant 2 T32 NS07091-06Al 0021.

PY - 1986/6

Y1 - 1986/6

N2 - The effects of enzymatic digestion, sodium borohydride reduction, acids used in décalcification procedures and techniques for inactivation of endogenous peroxidase were sequentially evaluated for their effect on the immunoreactivity of canine distemper virus in aldehyde-fixed paraffin-embedded tissue. Enzyme digestion improved immunoreactivity while sodium borohydride reduced background staining. Paraformaldehyde-glutaraldehyde-fixed tissues required thioglycolic acid treatment prior to enzyme digestion and sodium borohydride reduction to obtain results comparable to results obtained in formalin-fixed tissues. Detailed protocols for indirect immunofluorescence and the avidin-biotin-peroxidase complex procedure are provided.

AB - The effects of enzymatic digestion, sodium borohydride reduction, acids used in décalcification procedures and techniques for inactivation of endogenous peroxidase were sequentially evaluated for their effect on the immunoreactivity of canine distemper virus in aldehyde-fixed paraffin-embedded tissue. Enzyme digestion improved immunoreactivity while sodium borohydride reduced background staining. Paraformaldehyde-glutaraldehyde-fixed tissues required thioglycolic acid treatment prior to enzyme digestion and sodium borohydride reduction to obtain results comparable to results obtained in formalin-fixed tissues. Detailed protocols for indirect immunofluorescence and the avidin-biotin-peroxidase complex procedure are provided.

KW - aldehyde fixation

KW - canine distemper virus

KW - immunocytochemistry

KW - immunofluorescence technique

KW - immunoperoxidase

KW - technique

UR - https://www.scopus.com/pages/publications/0022574301

UR - https://www.scopus.com/pages/publications/0022574301#tab=citedBy

U2 - 10.1016/0166-0934(86)90016-9

DO - 10.1016/0166-0934(86)90016-9

M3 - Article

C2 - 3090088

AN - SCOPUS:0022574301

SN - 0166-0934

VL - 13

SP - 215

EP - 229

JO - Journal of Virological Methods

JF - Journal of Virological Methods

IS - 3

ER -

Immunocytochemical methods for demonstrating canine distemper virus antigen in aldehyde-fixed paraffin-embedded tissue

Abstract

Funding

Keywords

ASJC Scopus subject areas

Access to Document

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