Preparation of protein samples Sample Clauses

Preparation of protein samples. Snap-frozen cell pellets were lysed by resuspension in 100 µL of either 1% Triton X- 100 in PBS or RIPA buffer (Xxxxxxx et al. 2006, see table 2.2) as appropriate for the procedure. Samples were transferred into labelled 1.5 mL Eppendorf tubes and vortexed to further assist lysis. The samples were centrifuged at 16,160 x g for 10 minutes using a microcentrifuge (1-14 Microfuge, Sigma Centrifuges, Wem, UK) before the supernatants were transferred to clean 1.5 mL Eppendorf tubes. Two dilutions of each sample (1:10 and 1:20) were prepared (20 µL of each), using 1% Triton X-100 in PBS or RIPA buffer, for protein analysis. The remaining samples were kept on ice until further processing as described in section 2.2.3.
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