Trichloroacetic acid

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Cells are removed by centrifugation at 8,000 × g for 10 min. The supernatants are retained and sterilized by filtration through a 0.22-μm-pore-size Millex GP filter (Millipore, Billerica, MA).	- 20 ml of culture supernatants are precipitated by the addition of 100% (wt/vol) trichloroacetic acid (TCA) at −20°C to a final TCA concentration of 10% (incubated on ice for 45 min before centrifugation at 4,500 × g for 45 min at 4°C). 3 ml of ice-cold acetone is added to the pellet (15min before centrifugation at 4,500 × g for 45 min). The pellet is air dried before resuspension in 130 μl of rehydration buffer.

Upstream tips
- Cells are removed by centrifugation at 8,000 × g for 10 min. The supernatants are retained and sterilized by filtration through a 0.22-μm-pore-size Millex GP filter (Millipore, Billerica, MA).
Protocol tips
- 20 ml of culture supernatants are precipitated by the addition of 100% (wt/vol) trichloroacetic acid (TCA) at −20°C to a final TCA concentration of 10% (incubated on ice for 45 min before centrifugation at 4,500 × g for 45 min at 4°C). 3 ml of ice-cold acetone is added to the pellet (15min before centrifugation at 4,500 × g for 45 min). The pellet is air dried before resuspension in 130 μl of rehydration buffer.

Publication protocol

C. difficile strains were grown at 37°C in TY medium to stationary phase (24 h). Cells were removed by centrifugation at 8,000 × g for 10 min. The supernatants were retained and sterilized by filtration through a 0.22-μm-pore-size Millex GP filter (Millipore, Billerica, MA). Next, 20 ml of culture supernatants was precipitated by the addition of 100% (wt/vol) trichloroacetic acid (TCA) at −20°C to a final TCA concentration of 10%. Solutions were incubated on ice for 45 min before centrifugation at 4,500 × g for 45 min at 4°C. The supernatants were discarded, and 3 ml of ice-cold acetone was added to the pellet. The sample was then held on ice for 15 min before centrifugation at 4,500 × g for 45 min. The supernatant was discarded, and the pellet was air dried before resuspension in 130 μl of rehydration buffer (7 M urea, 2 M thiourea, 4% CHAPS {3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate}, 1% DTT, 0.5% ASB-14 detergent in Milli-Q water). Protein concentrations were determined using a Bradford protein assay (Bio-Rad, Mississauga, ON, Canada). A standard curve was generated using a bovine serum albumin (BSA) standard kit (Bio-Rad, Mississauga, ON, Canada).

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Protein isolation Bacteria - Clostridium difficile using Trichloroacetic acid from Sigma-Aldrich.

Manufacturer protocol

Download the product protocol from Sigma-Aldrich for Trichloroacetic acid below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Protein isolation Bacteria - Clostridium difficile using Trichloroacetic acid from Sigma-Aldrich. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.