Presence in bovine fetal serum of the protein antigenically related to p65-tumor associated antigen: its isolation and polyclonal antibody production.
Monoclonal antibodies raised to the 65-kDa tumor-associated protein (p65) isolated from a human breast cancer cell line have been used to detect an antigenically related protein (p65-like) present in fetal bovine serum (FBS) by Western blot analysis. We have isolated the p65-like protein from FBS by isoelectrofocusing (IEF) on native gels followed by electrophoresis in 12.5% polyacrylamide gel containing 0.1% SDS (SDS-PAGE). Immunostaining with anti-p65 monoclonal antibody of fetal bovine serum fractions separated by electrophoresis on cellulose acetate membrane revealed that the p65-like protein had a location similar to one of gamma-globulin. This protein migrates as a single band upon electrophoresis in SDS-PAGE and had four isoforms which migrate as two doublets with pI's of approximately 5.0 and 5.3.
Mirowski
M
M
Department of Biochemistry, Medical University, Lódøz, Poland.
Walaszek
Z
Z
Hanausek
M
M
D000818:Animals; D000911:Antibodies, Monoclonal; D000917:Antibody Formation; D000951:Antigens, Neoplasm / Q000097:blood* / Q000302:isolation & purification; D015153:Blotting, Western; D002417:Cattle; D003598:Cytoskeletal Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007525:Isoelectric Focusing; D007536:Isomerism; D008840:Microfilament Proteins; D010750:Phosphoproteins / Q000097:blood* / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D003598:Cytoskeletal Proteins; D008840:Microfilament Proteins; D010750:Phosphoproteins; C066559:Lcp1 protein, mouse