Cell-cell adhesion in the embryonic chick: Partial purification of liver adhesion molecules from liver membranes |
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Authors: | Larry D. Nielsen Malcolm Pitts Sharon R. Grady Edward J. McGuire |
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Affiliation: | 1. Division of Molecular and Cellular Biology, National Jewish Hospital and Research Center, 3800 E. Colfax Avenue, Denver, Colorado 80206 USA;2. Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, Colorado 80302 USA |
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Abstract: | Polyspecific xenoantisera specific for embryonic chick liver cells have been used to develop an assay for the detection of embryonic chick liver adhesion molecules (LAM). The neutralization of the antiaggregation immune sera and subsequent semiquantitative assessment of the remaining activity is the basis for this assay. Embryonic chick liver membrane proteins have been solubilized using a number of detergents and the LAM activity has been partially purified using sucrose-gradient sedimentation, isoelectric focusing, gel filtration, affinity chromatography, and preparative polyacrylamide gel electrophoresis procedures. The isolated LAM appears to be an intrinsic, trypsin-resistant, membrane glycoprotein of approximately 65,000 daltons. The 65K preparation of LAM is an effective immunogen in generating an aggregation blocking xenoantiserum specific for embryonic chick liver cells. Purification of LAM to homogeneity has been hampered by an apparent lability by self-association or loss by adsorption of the more purified LAM antigen(s). |
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Keywords: | To whom correspondence should be addressed. |
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