Fig 1.
Binding of PS K196E mutation-specific monoclonal antibodies to recombinant PS proteins.
The 96-well plates were coated with EGF-PS-K or EGF-PS-E (A) or FL-PS-K or FL-PS-E (B). The plates were then incubated with each of three monoclonal antibodies, 4B1, 15C8, and 16E3. HRP-labeled anti-mouse IgG antibody was used for the second antibody. Bound HRP was developed with TMB substrate, and the absorbance was measured at 450 nm. Error bars: SD (n = 3).
Fig 2.
Western blot analysis of the recombinant PS proteins using the PS K196E mutation-specific monoclonal antibody.
FL-PS-K and FL-PS-E were separated by SDS-PAGE under reducing conditions, transferred to a membrane, and incubated with the anti-His tag antibody (A) or the PS K196E mutation-specific monoclonal antibody 15C8 (B), and then with HRP-labeled anti-mouse IgG. The proteins were visualized using a chemiluminescent substrate. The molecular weights are shown on the left.
Fig 3.
Detection of PS K196E mutant in plasma by the PS K196E ELISA system.
Plasma samples diluted 20-fold using TBS were measured by the PS K196E ELISA system. The absorbance ranges of 122 KK and 11 KE plasma samples were −0.01–0.07 and 0.30–1.00, respectively. All data points represent the mean of three wells performed in triplicate.
Fig 4.
Dilution plots of FL-PS-E in the PS K196E ELISA system.
FL-PS-E was serially diluted from 0 μg mL−1 to 2.5 μg mL−1 in TBS (▲), 20-fold diluted PS-deficient plasma (+), or a 20-fold diluted KK plasma sample (■).