Linked Life Data

2449492

Source:http://linkedlifedata.com/resource/pubmed/id/2449492

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
3
pubmed:dateCreated
1988-4-1
pubmed:abstractText
We describe a technique for analysis by light microscopic immunocytochemistry of the distribution of plasmalemmal proteins in polarized epithelial cells. For this purpose, Madin Darby Canine Kidney (MDCK) cells were grown to confluency on Cytodex beads, the beads were fixed with formaldehyde, and semi-thin (0.5 micron) sections were cut at liquid nitrogen temperature on an ultracryomicrotome. The distribution of the basolaterally distributed plasmalemmal protein, Na,K-ATPase, was assessed by indirect immunofluorescence using a monospecific polyclonal antibody directed against the alpha-subunit of the Na pump. Such preparations enable epithelial monolayers to be evaluated in cross-section, thus permitting unambiguous topological assessment of apical and basolateral membrane proteins. Thus, the spatial uncertainties encountered in en face examination of membrane protein distribution in epithelia grown on solid supports are largely obviated. In addition, we describe a technique for removal of the bead matrix, which markedly reduces nonspecific background staining and improves access of reagents to the basal cell surface, thus permitting localization of basal lamina components.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Mar
pubmed:issn
0022-1554
pubmed:author
pubmed:issnType
Print
pubmed:volume
36
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
311-6
pubmed:dateRevised
2007-11-15
pubmed:meshHeading
pubmed:year
1988
pubmed:articleTitle
Immunocytochemical localization of plasmalemmal proteins in semi-thin sections of epithelial monolayers.
pubmed:affiliation
Yale University School of Medicine, Department of Cell Biology, New Haven, Connecticut 06510.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.