Comparative Physiology @ Guelph
Cellular Lights GFP-Tubulin Transduction on Silicon Strips
Protocol: Cellular Lights GFP-Tubulin Transduction on Silicon Strips
- Prepare Cells
- D-PBS w/o Ca2+ and Mg2+
- Cellular Lights GFP-Tubulin reagent
- BacMam Enhancer
- Dimethylsulfoxide (DMSO)
- Compatible Cell Line (see manufacturers list)
- Silicon Strip with 1 cm2 well
- Seed cells in silicon wells ~24 hours prior to transduction at a density that results in a 70-80% confluent layer
- Prepare 250 μL of Cellular Lights transduction solution by combining 90 μL of Cellular Lights reagent with 160 μL of D-PBS
- Aspirate regular growth medium from the prepared cells and add diluted Cellular Lights transduction solution to the cells. Incubate the cells at room temperature in the dark for 2-4 hours with gentle rocking or shaking.
- Aspirate Cellular Lights transduction solution from the plated cells and replace with appropriate culture medium with 1X BacMam Enhancer (1μL/mL). Incubate for 2 hours at 37°C and 5% CO2.
- Aspirate enhancer solution and replace with regular cell growth medium. Incubate cells for >16 hours to allow for expression of the protein.
- Transfer silicon strip to custom cell stretching device and mount on microscope.
Based on manufacturer’s protocol found at:
Written by Zac Robinson, Dec 2010