Antibodies to transforming growth factor-beta 2 peptides: specific detection of TGF-beta 2 in immunoassays.
Polyclonal antibodies have been raised to synthetic peptides corresponding to several regions of transforming growth factor-beta 2 (TGF-beta 2). All antisera were tested for their ability to react with either the native or reduced forms of both TGF-beta 1 and TGF-beta 2 in enzyme-linked immunosorbent assays, Western blots, and immunoprecipitation assays. On Western blots, antisera raised to a peptide corresponding to residues 50-75 of TGF-beta 2 specifically detected 5 ng TGF-beta 2, while antisera raised to regions 1-30 and 79-108 cross-reacted with TGF-beta 1. Anti-P 50-75(2) also localized TGF-beta 2 in murine placenta in immunohistochemical studies. In immunoprecipitation assays with either iodinated TGF-beta s or with media conditioned by cells labeled with [35S]cysteine, both anti-P 50-75(2) and anti-P 79-108(2) specifically immunoprecipitated TGF-beta 2 under reducing conditions only, while anti-P 79-108(2) also reacted with TGF-beta 1. None of the TGF-beta 2 peptide antibodies was able to block receptor binding of either TGF-beta 1 or 2. Analysis of the cross-reactivity patterns of these peptide antibodies suggests conformational differences between TGF-beta 1 and TGF-beta 2.
Flanders
K C
KC
Laboratory of Chemoprevention, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892.
Cissel
D S
DS
Mullen
L T
LT
Danielpour
D
D
Sporn
M B
MB
Roberts
A B
AB
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D001667:Binding, Competitive; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D051379:Mice; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology; D010920:Placenta / Q000378:metabolism; D011247:Pregnancy; D015291:Transforming Growth Factors / Q000032:analysis / Q000276:immunology* / Q000378:metabolism
D000906:Antibodies; D010446:Peptide Fragments; D015291:Transforming Growth Factors