TY - JOUR
T1 - Accurate quantitation of native Gc in serum and estimation of endogenous Gc
T2 - G-actin complexes by rocket immunoelectrophoresis
AU - Goldschmidt-Clermont, Pascal J.
AU - Galbraith, Robert M.
AU - Emerson, David L.
AU - Werner, Philip A M
AU - Nel, Andre E.
AU - Lee, William M.
N1 - Funding Information:
Publication Number 722 from the Department of Basic and Clinical Immunology and Microbiology, Medical University of South Carolina. This research was supported in part by NIH Grant CA-27062, Institutional Awards CR17 and GR45, and Labcatal Laboratories, Paris, France. R.M.G. was the recipient of NIH Research Career Development Award CA-00611 and P.J.G.-C. was supported by a Clinical Post-doctoral Fellowship from the Medical University of South Carolina. Our thanks are due to Phillippe Arnaud for critical review of this manuscript.
PY - 1985/6/14
Y1 - 1985/6/14
N2 - Complex formation between purified Gc and G-actin caused increased rocket height on immunoelectrophoresis with monospecific Gc antiserum, and artefactually high calculated Gc levels. The increase in rocket height varied in log:linear fashion with the amount of G-actin present, up to a plateau attained at equimolarity. The raw Gc values could therefore be corrected to within ± 10% of known levels by addition of excess G-actin and use of standard plots obtained with Gc after saturation with G-actin. This also allowed quantitation of the percentage of Gc complexed with G-actin. In subsequent studies of whole human sera, comparison of normal controls with pregnant subjects and patients with liver disease showed evidence of differences both in absolute quantities of Gc and the relative proportion circulating as complex with G-actin. This appeared to be due to increased release of cellular actin into the extracellular space. These results show that rocket immunoelectrophoresis can be modified to provide accurate Gc levels, and also information concerning different molecular forms of this protein.
AB - Complex formation between purified Gc and G-actin caused increased rocket height on immunoelectrophoresis with monospecific Gc antiserum, and artefactually high calculated Gc levels. The increase in rocket height varied in log:linear fashion with the amount of G-actin present, up to a plateau attained at equimolarity. The raw Gc values could therefore be corrected to within ± 10% of known levels by addition of excess G-actin and use of standard plots obtained with Gc after saturation with G-actin. This also allowed quantitation of the percentage of Gc complexed with G-actin. In subsequent studies of whole human sera, comparison of normal controls with pregnant subjects and patients with liver disease showed evidence of differences both in absolute quantities of Gc and the relative proportion circulating as complex with G-actin. This appeared to be due to increased release of cellular actin into the extracellular space. These results show that rocket immunoelectrophoresis can be modified to provide accurate Gc levels, and also information concerning different molecular forms of this protein.
KW - G-actin complexes
KW - Gc
KW - Rocket immunoelectrophoresis
KW - Vitamin D3
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U2 - 10.1016/0009-8981(85)90144-5
DO - 10.1016/0009-8981(85)90144-5
M3 - Article
C2 - 4042352
AN - SCOPUS:84886639954
SN - 0009-8981
VL - 148
SP - 173
EP - 183
JO - Clinica Chimica Acta
JF - Clinica Chimica Acta
IS - 3
ER -