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Isolation of specific Rab-binding proteins from cell lysates by GST pull-down assay

Sackmann-Sala, L., Ding, J., Frohman, L

Until a disease-modifying therapy is found, symptomatic treatments, including a drug for constipation, remain under development

Cells were collected in PBS and sequentially lysed in buffer 1 (50 mM HEPES pH 7.5, 140 mM NaCl, 1 mM EDTA, 10% glycerol, 0.5% NP-40, 0.25% TritonX-100), buffer 2 (10 mM Tris pH 8, 200 mM NaCl, 1 mM EDTA, 0.5 mM EGTA) and buffer 3 (10 mM Tris pH 8, 100 mM NaCl, 1 mM EDTA, 0.5 mM EGTA, 0.1% sodium deoxycholate, 0.5% N-lauroylsarcosine) with protease inhibitors (Roche)

PMID: 27358198

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