Characterization of two preparations of antibodies to basic fibroblast growth factor which exhibit distinct patterns of immunolocalization.
Immunoglobulins reactive against basic fibroblast growth factor (bFGF) were obtained from the serum of a single rabbit immunized against residues [1-24] of bFGF conjugated to keyhole limpet hemocyanin (KLH). Pure immunoglobulin preparations no. 1 and no. 2 were prepared using different affinity chromatography columns and preabsorption to KLH-coupled Sepharose for preparation no. 1. Both preparations no. 1 and no. 2 were specific for bFGF in in vitro assays. Competition with synthetic peptides suggests that preparations no. 1 and no. 2 recognize predominantly epitope(s) within residues [16-24]bFGF or residues [1-10]bFGF, respectively, in situ. Furthermore, no. 2 (but not no. 1) antibodies can react with tissue-(heparin-)-bound antigen. When used in indirect immunofluorescence for bFGF in frozen heart sections, preparation no. 1 stained predominantly muscle intercalated discs (IcDs); muscle nuclei were also stained, in an overall punctate fashion. Preparation no. 2 stained muscle nuclei strongly, in association with the nuclear envelope; it also stained basement-membrane associated bFGF. Differences in immunostaining were also observed in uterine smooth muscle and kidney sections but not in skeletal muscle. It is plausible that accessibility of various epitopes within the amino-terminal region depends strongly on the local interactions of bFGF. Our data illustrate the importance of using several different antibodies to localize bFGF in a tissue.
Kardami
E
E
St Boniface General Hospital Research Centre, University of Manitoba, Winnipeg, Canada.
Murphy
L J
LJ
Liu
L
L
Padua
R R
RR
Fandrich
R R
RR
D000042:Absorption; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D001667:Binding, Competitive; D015153:Blotting, Western; D002417:Cattle; D002846:Chromatography, Affinity; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D016222:Fibroblast Growth Factor 2 / Q000276:immunology* / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006493:Heparin / Q000494:pharmacology; D009206:Myocardium / Q000378:metabolism*; D011817:Rabbits; D051381:Rats
D000906:Antibodies; D000939:Epitopes; D016222:Fibroblast Growth Factor 2; D006493:Heparin