Immunoblotting studies of the molecular forms of protein C in plasma.
Anti-plasma protein C monoclonal antibodies were prepared and characterized, and quantitative immunoblotting techniques were developed to determine the molecular forms of protein C in whole plasma. Two antibodies reacted with the heavy chain of protein C, four reacted with the light chain, and two reacted only with nonreduced protein C. A doublet of protein C (MW = 63-66K) was seen on nonreduced immunoblots of normal plasma and 30 heterozygous protein C deficient plasmas (2-77% protein C antigen). In reduced plasma, approximately 75% of protein C presented as doublet heavy chains (MW = 39-42K) and doublet light chains (MW = 22-25K), and approximately 25% was single chain (MW = 64K). The immunoblotting technique was quantitative, specific, sensitive, and correlated with electroimmunoassay results. It also provided visual qualitative information not obtainable with other methods of quantitation.
Heeb
M J
MJ
Department of Immunology, Research Institute of Scripps Clinic, La Jolla, CA 92037.
Schwarz
H P
HP
White
T
T
Lämmle
B
B
Berrettini
M
M
Griffin
J H
JH
D000911:Antibodies, Monoclonal; D006801:Humans; D007118:Immunoassay; D015151:Immunoblotting / Q000379:methods*; D008970:Molecular Weight; D011486:Protein C / Q000276:immunology / Q000302:isolation & purification*; D020151:Protein C Deficiency; D011487:Protein Conformation
D000911:Antibodies, Monoclonal; D011486:Protein C