Trichloroacetic acid

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- The supernatants are precipitated with 10% trichloroacetic acid. For concentration by desalinization, washed with acetone, and then dried at room temperature. The dried pellets and the remaining cells were suspended in a reducing sample buffer containing 2-mercaptoethanol.

Protocol tips
- The supernatants are precipitated with 10% trichloroacetic acid. For concentration by desalinization, washed with acetone, and then dried at room temperature. The dried pellets and the remaining cells were suspended in a reducing sample buffer containing 2-mercaptoethanol.

Publication protocol

The supernatants were precipitated with 10% trichloroacetic acid (Sigma) for concentration by desalinization, washed with acetone, and then dried at room temperature. The dried pellets and the remaining cells were suspended in a reducing sample buffer containing 2-mercaptoethanol. Samples were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Separated proteins were electrotransferred to polyvinylidene difluoride membranes (Millipore, Bedford, MA) with a semidry electroblotter (Bio-Rad, Richmond, CA). Membranes were blocked with 1% (wt/vol) skimmed milk in Tris-buffered saline (TBS, pH 7.5) containing 0.05% Tween 20 (TBS-T) and incubated with mouse anti-IL-1β antibody (Cell Signaling Technology, Beverly, MA) or mouse anti-α-tubulin antibody (Cedarlane, Ontario, Canada) in Immuno-enhancer reagent A (Wako Pure Chemical Industries) overnight at 4°C. After being washed with TBS-T, membranes were incubated with a horseradish peroxidase-conjugated goat anti-mouse IgG antibody (Sigma) in Immuno-enhancer reagent B for 1 h at room temperature. Labeled proteins were visualized with ImmunoStar LD Western blotting substrate (Wako Pure Chemical Industries) or Western blotting substrate (Thermo Scientific, Waltham, MA).

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 Mammalian cells - THP-1 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 Mammalian cells - THP-1 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.