This protocol describes how to use Carbotrace 680 to determine cellulose content in a sample.
Solutions and Reagents:
Carbotrace 680 is provided as concentrated solution. The following common reagents are required (not supplied):
- Phosphate buffered saline (PBS), pH 7.4
- 96-well plate (round bottom)
- Spectrophotometer
Assay Procedure:
- Prepare a dilution series of your cellulose containing sample im PBS (e.g.: 1, 1:2, 1:5, 1:10, 1:100, 0)
- Dilute Carbotrace 680 in PBS 1:500.
- Add 50 µl of each cellulose containing dilution into a well of a 96-well plate.
- Add 50 µl diluted Carbotrace 680 to each cellulose containing dilution and the blank control.
- Measure emission using spectrophotometer settings (see 'Spectrophotometer Settings' below).
Spectrophotometer Settings:
- Carbotrace 680: Excite at 540 nm and collect emission at 680 nm. Optional: Record an emission spectrum (560 - 800 nm) with 540 nm excitation.