- 0.15% (w/v) sodium deoxycholate
- 72% (w/v) Trichloroacetic acid (TCA)
- Copper tartrate/carbonate (CTC)
- 20% (v/v) Folin-Ciocalteu reagent
- Bovine serum albumin (BSA)
- Spectrophotometer and tubes
- Prepare standard dilutions of BSA of 25, 50, 75, and 100 mg/mL. Prepare
appropriate serial dilutions of the sample to be measured.
- Place 1.0 mL of each of the above into separate tubes. Add 100 mL of
sodium deoxycholate to each tube.
- Wait 10 minutes and add 100 mL of TCA to each tube.
- Centrifuge each tube for 15 minutes at 3000 xg and discard the supernatant.
- Add 1.0 mL of water to each tube to dissolve the pellet. Add 1.0 mL of
water to a new tube to be used as a blank.
- Add 1.0 mL of CTC to each tube (including the blank), vortex and allow
to set for 10 minutes.
- Add 500 mL Folin-Ciocalteu to each tube, vortex and allow to set for 30
- Turn on and zero a spectrophotometer to a wavelength of 750 nm. Use the
blank from step 7 to adjust for 100% T.
- Read each of the standards and samples at 750 nm.
- Plot the absorbance of the standards vs their concentration. Compute the
extinction coefficient and calculate the concentrations of the unknown