Figure 1. Characterization of specific antibodies against OPN-FL, OPN-R and OPN-L and development of specific ELISAs.
Recombinant human OPN-FL was digested with thrombin (IIa) or thrombin (IIa)/CPB to produce OPN-R and OPN-L respectively. A. Western blot analysis of human OPN-FL, OPN-R and OPN-L with the specific anti-peptide antibodies anti-OPN, anti-OPN-R and anti-OPN-L. OPN-R (SLAYGLR) and OPN-L (SVVYGL) peptides were used for competitive inhibition of antibody binding. B, ELISAs for OPN-FL (commercial), OPN-R and OPN-L ELISA. C. Competition of substituted peptides for binding of anti-OPN-R and anti-OPN-L to immobilized purified OPN-R and OPN-L respectively. Inhibition of binding by the cognate unsubstituted peptide was defined as 100%.
