Tethered Particle Motion - Chip and Tether Assembly - Tether Assembly

Tether Assembly

First, the chip has to be passivated so that the polymer won't stick to the glass, there are plenty of blocking reagents available (BSA, alpha-casein, etc.) and one should find what works best for the specific situation Next, the surface should be coated with an antibody or other reactive molecule (such as anti-digoxigenin) that will bind to an antigen (digoxigenin) at one end of the polymer. After an incubation of about 45min, the excess antibody has to be washed away. After washing the excess antibody, the polymer should be injected into the chip and incubated for about the same time. The polymer had been modified before at the ends. One end has a biotin tail and the other has a digoxigenin tail. After incubation, unbound polymer has to be washed out from the cell. Then, anti-biotin coated beads should be injected to the flowcell and incubate for about 30-45min. Excess beads should be washed out.