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Western blot HEK-293T cells expressing hCC WT or the L68Q variant were washed twice with ice-cold phosphate-buffered saline (PBS) and lysed on ice using a freshly prepared ice-cold cell lysis buffer containing 50 mM Tris-HCl, pH 7.4, 100 mM NaCl, 50 mM -glycerophosphate, 10% glycerol (w/v), 1% NP-40 (w/v), 1 mM EDTA, 2 mM NaVO4, and a complete, EDTA-free, protein inhibitor cocktail (Roche Applied Science, Mannheim Germany) at 20 l per mL of lysis buffer

Gatto D, Paus D, Basten A, Mackay CR, Brink R

It is an 83-residue recombinant protein, made by expressing an engineered gene in Escherichia coli and purifying the product, not by solid-phase chemical synthesis

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