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PMID: 9072183 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Antigen retrieval in cryostat tissue sections and cultured cells by treatment with sodium dodecyl sulfate (SDS).

Histochemistry and cell biology ·Vol. 105 ·No. 4 ·1996-04-00 ·Pages 261-7

Brown D, Lydon J, McLaughlin M, Stuart-Tilley A, Tyszkowski R, Alper S

Abstract

A simple method for antigen retrieval in tissue sections and cell cultures is described. Because many antibodies recognize denatured proteins on western blots, but are poorly reactive by immunocytochemistry, the effect of applying sodium dodecyl sulfate (SDS) to cryostat sections of tissues and to cell cultures prior to immunostaining was examined. In many cases, a 5-min pretreatment with 1% SDS produced a dramatic increase in staining intensity by indirect immunofluorescence. Among the antibodies tested that showed a positive effect of SDS were an anti-Na/K-ATPase monoclonal antibody, an anti-AE1/2 anion exchanger polyclonal antipeptide antibody, a monoclonal anti-caveolin antibody, and an anti-rab4 monoclonal antibody. In other cases, including antibodies against gp330, aquaporin 1, and aquaporin 2, no effect of SDS was detected. The results show that SDS treatment can be used as a simple method of antigen retrieval in cryostat sections and on cultured cells. In some cases, antigens were not detectable without pretreatment with SDS.

MeSH Terms
Animals Antigens/analysis Cell Line Cryoultramicrotomy Epithelial Cells Epithelium/chemistry Immunohistochemistry/methods Kidney/chemistry,cytology Rats Rats, Sprague-Dawley Sodium Dodecyl Sulfate/chemistry
Chemicals
Antigens Sodium Dodecyl Sulfate
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Brown D
Department of Pathology, Massachusetts General Hospital, Boston, MA 02129, USA.
Lydon J
McLaughlin M
Stuart-Tilley A
Tyszkowski R
Alper S
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Article Info
Journal
Histochemistry and cell biology
Abbr.
Histochem Cell Biol
ISSN
0948-6143
Published
1996-04-00
Pages
261-7
Language
English
Region
Germany
NLM ID
9506663
Subset
IM
Grants
NIDDK NIH HHS · DK 42956 · United States
NIDDK NIH HHS · DK 43495 · United States
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