To achieve signal amplification, streptavidin was attached via biotin to the second antibody.
The second antibody (HRP conjugate, YEASON) was 1 5,000 diluted and incubated for 1 h at room temperature.
The second antibody (goat against rabbit Fc portion of Ig) was a product of Sigma-Aldrich (USA).
With property of good conductivity and large surface area, porous PtPd nanochains can conjugate more GQDs and second antibody to improve the sensitivity of the immunoassay.
The use of a second antibody against N1 offered much greater specificity and reliability than the previous biosensor protocol.
One of the antibodies was coated on magnetic microparticles, and the second antibody was conjugated with a reporter group.
Upon the formation of immunocomplex, the second antibody labeled with horseradish peroxidase was simply immobilized on the electrode surface.
The primary antibody and second antibody were sequentially added.
As a second antibody, Alexa Fluor 555 goat anti-rat IgG (Invitrogen, Carlsbad, CA) was used.
After incubation with the relative second antibody, immune complexes were detected using the ECL method.
This was confirmed using a second antibody specific for the beta7 integrin chain alone.
