Competitive enzyme immunoassay for quantification of the cellular form of fibronectin (EDAcFN) in blood samples.
A competitive enzyme immunoassay based on the monoclonal antibody DH1 was developed for the quantification of extradomain A (EDA)-containing isoform of cellular fibronectin (cFN). The average EDAcFN concentration in normal plasma was 2.46 micrograms/ml and in serum 0.68 micrograms/ml. Similar results were obtained by gelatin-Sepharose binding and immunoblotting. Studies on the effect of storage of plasma and serum as frozen samples indicated that the amount of EDAcFN decreased rapidly when stored at -20 degrees C. Storage at -70 degrees C resulted in less of a decrease in the concentration of EDAcFN. The method permits the determination of EDAcFN in blood samples from diseases showing increased amounts of the protein at the tissue level.
Ylätupa
S
S
Department of Anatomy, University of Helsinki, Finland.
Partanen
P
P
Haglund
C
C
Virtanen
I
I
D000911:Antibodies, Monoclonal; D001773:Blood Cells / Q000737:chemistry; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005353:Fibronectins / Q000097:blood*; D006801:Humans; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D011489:Protein Denaturation; D015203:Reproducibility of Results
D000911:Antibodies, Monoclonal; D005353:Fibronectins