Triton X 100 1 L

Triton X-100

(TX-100; octoxynol 9; 4-(1,1,3,3-Tetramethylbutyl)phenyl-polyethylene glycol, t-Octylphenoxypolyethoxyethanol, Octylphenoxypolyethoxyethanol, Polyethylene glycol tert-octylphenyl ether)

Triton X-100 is a non-ionic detergent that is suitable for solubilizing membrane proteins.

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Protocol: Triton X-100 for the Recovery Inclusion Bodies

1) Centrifuge the cell lysate at maximum speed for 15 minutes at 4 degrees Celsius in a microcentrifuge.

2) Decant the supernatant.  Resuspend the pellet in 9 volumes of Cell lysis buffer (50 mM Tris HCl pH 8.0, 10 mM EDTA (pH 8.0), 100 mM NaCl, 0.5 % Triton X-100) on ice.

3) Store the suspension for 5 minutes at room temperature.

4) Centrifuge the tube at maximum speed for 15 minutes at 4 degrees in a microfuge.

5) Decant the supernatant and set it aside for the next step.  Resuspend the pellet in 100 ul of H₂O.

6)  Remove 10 ul samples of the supernatant and of the reuspended pellet.  Mix each sample with 10 ul of 2x SDS gel loading buffer and analyze the samples of SDS-PAGE to determine which fraction contains the protein of interest.