Linked Life Data

19378059

Source:http://linkedlifedata.com/resource/pubmed/id/19378059

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:dateCreated
2009-4-20
pubmed:abstractText
A method for the affinity purification of autoantibodies from small volumes of human sera using a western blot strip containing a target antigen electrophoretically transferred from a sodium dodecyl sulfate (SDS) polyacrylamide gel is described. This method is a very useful alternative for affinity column chromatography, particularly when the antigen of interest is of low abundance. The protein mixture is resolved on a preparative SDS polyacrylamide gel and transferred to nitrocellulose membrane. A couple of strips are excised vertically from either side of the blotted membrane and immunoblotted with specific antisera to identify the target band. Then the target band is excised horizontally and used for affinity purification. We have used this procedure to affinity purify antibodies to a 70,000 molecular weight protein derived from HeLa cell extract. A sham band, excised away from the target antigen, was used as a control for sham purification of autoantibodies. The autoantibodies purified in this manner reproduced the multiple nuclear dot anti-nuclear antibody pattern obtained using crude sera from 21 patients without primary biliary cirrhosis or anti-mitochondrial antibody.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:issn
1064-3745
pubmed:author
pubmed:issnType
Print
pubmed:volume
536
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
201-11
pubmed:meshHeading
pubmed:year
2009
pubmed:articleTitle
Affinity purification of autoantibodies from an antigen strip excised from a nitrocellulose protein blot.
pubmed:affiliation
Oklahoma Medical Research Foundation, 825 NE 13th Street, OKC, OK, 73104, USA. biji-kurien@omrf.org
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't, Research Support, N.I.H., Extramural