Characterization of antisera directed against follistatin/activin-binding protein peptides.
An attempt was made to develop immunologic probes directed against follistatin/activin-binding protein (ABP), for use in investigating the distribution of ABP in various tissues. Five oligopeptides corresponding to different regions of the predicted ABP amino acid sequence (peptides 1-12, 28-43, 123-134, 270-281 and 300-315) were synthesized chemically, and coupled to Limulus polyphemus hemolymph hemocyanin. The peptide-hemocyanin conjugates were then used to immunize rabbits, and the immunoresponses were monitored by enzyme-linked immunosorbent assay. Reactivity of the antipeptide antisera with ABP was determined by SDS-polyacrylamide gel electrophoresis and immunoblotting analysis. All of the peptides produced immune responses. The antiserum to peptide 123-134 recognized all forms of ABP, whereas the antiserum to peptide 300-315 reacted specifically and sensitively with the long forms of ABP. These two antisera exhibited only a limited cross-reaction with other proteins or none at all. Therefore, they will be useful for studying the distribution of ABP in various tissues.
Saito
S
S
Frontier Research Program, Institute of Physical and Chemical Research (RIKEN), Saitama, Japan.
Sugino
K
K
Yamanouchi
K
K
Kogawa
K
K
Titani
K
K
Shiota
K
K
Takahashi
M
M
Sugino
H
H
D000595:Amino Acid Sequence; D000818:Animals; D000917:Antibody Formation; D000918:Antibody Specificity; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D038681:Follistatin; D006023:Glycoproteins / Q000276:immunology*; D007106:Immune Sera / Q000276:immunology*; D007114:Immunization; D008969:Molecular Sequence Data; D011817:Rabbits
D038681:Follistatin; D006023:Glycoproteins; D007106:Immune Sera