PMID: 7538416Feb 1, 1995Paper

Expression of the alternatively spliced EIIIB segment of fibronectin

Cell Adhesion and Communication
J H PetersRichard O Hynes

Abstract

Previous descriptions of the expression and distribution of the alternatively spliced EIIIB segment of fibronectin (FN) relied upon an antibody which, on subsequent testing, was shown not to recognize this segment directly. This raises concerns regarding the reliability of all such previous descriptions. In order to prepare reagents directly reactive with this segment, we raised polyclonal antibodies to two different bacterial fusion proteins containing intact EIIIB segments, and to a synthetic 36 amino acid peptide from the center of the EIIIB segment. Antibodies raised to each of these three immunogens recognized fusion proteins containing the EIIIB segment, but failed to recognize full length EIIIB+ FNs produced by mammalian cells, suggesting that oligosaccharide linked to Asn1359 within the EIIIB segment, or potentially to other residues in FN, might interfere with antibody recognition of this segment. Consistent with this hypothesis, N-deglycosylation of recombinant full and partial length EIIIB+ FNs permitted their specific recognition by the anti-fusion protein (but not anti-peptide) antibodies. Using anti-fusion protein antibodies coupled with deglycosylation procedures, we provide a series of new results relevant to th...Continue Reading

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