ISOLATION AND SEPARATION OF HUMAN CYTOKERATINS USING VARIOUS CHROMATOGRAPHIC TECHNIQUES

Persistent Link:
http://hdl.handle.net/10150/276475
Title:
ISOLATION AND SEPARATION OF HUMAN CYTOKERATINS USING VARIOUS CHROMATOGRAPHIC TECHNIQUES
Author:
Meiklejohn, Bruce Ian, 1959-
Issue Date:
1987
Publisher:
The University of Arizona.
Rights:
Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
Abstract:
The cytokeratins from various human tissue were isolated using chromatographic techniques. The cytokeratins were first extracted from crude tissue using high and low salt buffers. It was necessary to use a denaturing agent such as urea to solubilize the resulting cytokeratin pellet. Imidazole also seemed to help solubilize the pellet and a reducing agent such as 2-Mercaptoethanol was not needed as previously believed. The acidic cytokeratins were separated from the neutral-basic cytokeratins using a DEAE ion-exchange column. The acidic cytokeratin fraction was further separated on a moderately polar reverse phase column with an acetonitrile gradient to eluted the proteins. Tetramethylammonium tetrafluoroborate was added to the mobile phase to react with any unreacted silanol groups on the stationary phase and trifluoroacetic acid was added to ion pair with the protein. The peaks were analyzed for purity using two dimensional electrophoresis and monoclonal antibodies that recognize the cytokeratins.
Type:
text; Thesis-Reproduction (electronic)
Keywords:
Cytoplasmic filaments.; Intermediate filament proteins.
Degree Name:
M.S.
Degree Level:
masters
Degree Program:
Graduate College; Chemistry
Degree Grantor:
University of Arizona

Full metadata record

DC FieldValue Language
dc.language.isoen_USen_US
dc.titleISOLATION AND SEPARATION OF HUMAN CYTOKERATINS USING VARIOUS CHROMATOGRAPHIC TECHNIQUESen_US
dc.creatorMeiklejohn, Bruce Ian, 1959-en_US
dc.contributor.authorMeiklejohn, Bruce Ian, 1959-en_US
dc.date.issued1987en_US
dc.publisherThe University of Arizona.en_US
dc.rightsCopyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.en_US
dc.description.abstractThe cytokeratins from various human tissue were isolated using chromatographic techniques. The cytokeratins were first extracted from crude tissue using high and low salt buffers. It was necessary to use a denaturing agent such as urea to solubilize the resulting cytokeratin pellet. Imidazole also seemed to help solubilize the pellet and a reducing agent such as 2-Mercaptoethanol was not needed as previously believed. The acidic cytokeratins were separated from the neutral-basic cytokeratins using a DEAE ion-exchange column. The acidic cytokeratin fraction was further separated on a moderately polar reverse phase column with an acetonitrile gradient to eluted the proteins. Tetramethylammonium tetrafluoroborate was added to the mobile phase to react with any unreacted silanol groups on the stationary phase and trifluoroacetic acid was added to ion pair with the protein. The peaks were analyzed for purity using two dimensional electrophoresis and monoclonal antibodies that recognize the cytokeratins.en_US
dc.typetexten_US
dc.typeThesis-Reproduction (electronic)en_US
dc.subjectCytoplasmic filaments.en_US
dc.subjectIntermediate filament proteins.en_US
thesis.degree.nameM.S.en_US
thesis.degree.levelmastersen_US
thesis.degree.disciplineGraduate Collegeen_US
thesis.degree.disciplineChemistryen_US
thesis.degree.grantorUniversity of Arizonaen_US
dc.identifier.proquest1331417en_US
dc.identifier.oclc18009086en_US
dc.identifier.bibrecord.b1837136xen_US
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