2. Coat with Biotin BSA
Infuse biotinylated BSA at 2 mg/ml into the flow cell until the entire volume is filled (~15 µl). Incubate for 10 minutes.
Wash the flow cell by wicking 60 µl of Wash Buffer through the chamber.
3. Add Streptavidin
Infuse 20 µl of streptavidin at 10 mg/ml. Incubate for 10 minutes.
Wash the flow cell by wicking 60 µl of Wash Buffer through the chamber.
4. Block with Pluronic
Infuse 20 µl of 1% Pluronic F-127. Incubate for 1 minute.
Wash the flow cell by wicking 60 µl of Wash Buffer through the chamber.
5. Add Microtubules
Infuse 20 µl of biotinylated microtubules at ~1 µM [tubulin]. Microtubules can be diluted in 1X Tubulin PEM Buffer if necessary. Incubate for 20 minutes.
Wash the flow cell by wicking 60 µl of Flow Cell Buffer through the chamber.
NOTE :
Flow Cell Buffer is used as the final wash because it contains the oxygen scavenging mix required for imaging. It is necessary to make fresh Flow Cell Buffer every couple of hours to maintain optimal performance of the Oxygen Scavenging Mix.