Antibodies against the cystic fibrosis transmembrane regulator.
Rabbit polyclonal antibodies have been raised against high-performance liquid chromatography purified synthetic peptides corresponding to two discrete regions of the cystic fibrosis transmembrane regulator (CFTR) protein: the R-domain (residues 785-796) and the extreme COOH-terminus (residues 1467-1480). Antibodies (Ab) to each of these peptides were affinity purified either by passage over a peptide-derivatized agarose matrix (Ab 785) to produce monospecific polyclonal antibodies or by protein A affinity chromatography to purify the immunoglobulin G1 fraction free of other serum proteins (Ab 1467). These antibodies recognize a candidate CFTR protein in the colonic cell line T84, as determined by Western blot analysis and by immunoprecipitation and labeling of the precipitate with [gamma-32P]ATP in the presence of protein kinase A. Both antibodies precipitated CFTR-related polypeptides from four mammalian cell types (HeLa, Bsc-40, HEp-2, and Chinese hamster ovary cells) transfected with the full-length CFTR cDNA clone using a vaccinia T7 protein expression system. Similar results were observed using a yeast CFTR expression system. In each case the Mr values of the bands observed were consistent with that expected for the CFTR protein. These antibodies should be useful probes for the immunocytochemical localization, immunoaffinity purification, and ultimately the functional characterization of the CFTR protein.
Department of Physiology and Biophysics, University of Alabama, Birmingham 35294-0005.
C M
CM
Fuller
M B
MB
Howard
D M
DM
Bedwell
R A
RA
Frizzell
D J
DJ
Benos
D000818:Animals; D000906:Antibodies / Q000276:immunology* / Q000302:isolation & purification; D015153:Blotting, Western; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003550:Cystic Fibrosis / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D006801:Humans; D008565:Membrane Proteins / Q000276:immunology*; D010455:Peptides / Q000302:isolation & purification; D011233:Precipitin Tests; D015003:Yeasts / Q000166:cytology / Q000378:metabolism
D000906:Antibodies; C505032:CFTR protein, human; D008565:Membrane Proteins; D010455:Peptides; D019005:Cystic Fibrosis Transmembrane Conductance Regulator