Immunohistochemical detection of cytokines in paraffin-embedded mouse tissues.
J Immunol Methods
; 210(1): 103-8, 1997 Dec 15.
Article
in En
| MEDLINE
| ID: mdl-9502589
ABSTRACT
We have successfully developed a method for the immunohistochemical detection of interleukin 2 (IL-2), IL-4, IL-6, IL-10O, IFNgamma and TNFalpha using monoclonal antibodies (MAb), in sections of mouse tissue embedded in paraffin wax. The method involved fixation in periodate-lysine-paraformaldehyde (PLP), rapid dehydration and infiltration under vacuum with paraffin wax at 54 degrees C. Comparative observations demonstrated that the method gives equivalent or better results than formaldehyde fixed, frozen sections. Since reliable controls, both positive and negative, are paramount for interpretation of immunohistochemical staining, such controls were determined. The following tissues were shown to be suitable as positive controls when using paraffin-embedding spleen for the detection of TNFalpha, small intestine for IL-2, IL-4 and IL-10, and HSV-1 infected eyes for IL-6 and IFNgamma. We conclude that PLP fixation and low temperature paraffin-embedding is a method which provides both preservation of excellent tissue morphology and reliable immunohistochemical identification of cytokines. These attributes will be invaluable in a wide variety of experimental situations.
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Cytokines
/
Immunoenzyme Techniques
Type of study:
Diagnostic_studies
Limits:
Animals
Language:
En
Journal:
J Immunol Methods
Year:
1997
Document type:
Article
Affiliation country:
United kingdom
Publication country:
HOLANDA
/
HOLLAND
/
NETHERLANDS
/
NL
/
PAISES BAJOS
/
THE NETHERLANDS