Production of oligoclonal antibodies directed to the N-terminal of smooth muscle alpha actin using peptidyl-polyacrylic resins as direct immunogens.
Polyacrylamide-based solid-phase supports were used in the production of a synthetic peptide analogous to amino acid sequence 1-9 from bovine aortic actin. Different peptidyl-resins were prepared and directly used to induce an antibody response in rabbits. Antisera that recognized the resin-bound peptide were compared. Some of them reacted with native aortic actin; this response was highly specific for the smooth muscle alpha actin isotype. We also immunized rabbits with the 1-9 peptide linked to bovine serum albumin. In this case, antisera were less reactive with native aortic actin, and cross-reacted with the two closely related smooth muscle alpha and macrophage actin isotypes. This method provides a rapid and simple procedure for generating peptide immunogens and antibodies to them, by the combination of peptidyl-resin immunizations and resin-bound peptide immunoassays.
Hanin
V
V
Centre de Recherches de Biochimie Macromoléculaire, UPR 8402 CNRS et U249 INSERM, Montpellier, France.
Naharisoa
H
H
Sarrade
V
V
Calas
B
B
D000199:Actins / Q000276:immunology*; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D000917:Antibody Formation; D001011:Aorta / Q000378:metabolism; D002417:Cattle; D003429:Cross Reactions; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000378:metabolism; D010455:Peptides / Q000138:chemical synthesis* / Q000276:immunology; D011817:Rabbits / Q000276:immunology; D012116:Resins, Plant; D012689:Sequence Homology, Nucleic Acid
D000199:Actins; D000906:Antibodies; D010455:Peptides; D012116:Resins, Plant