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Twenty microliters of supernatant was transferred into a 96-well plate, and 200 l of 37 C o-dianisidine hydrochloride solution was added immediately before the optical density was read at 450 nm and again 30 s later, as previously described 26

Understanding these differences is essential for designing precise and reproducible experiments

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The resin was packed into a glass column and washed with 20 column volumes of 20 mM HEPES pH 7.4, 100 mM NaCl, 2 mM MgCl 2 , 5 mM CaCl 2 , 1 M exendin-4 or 10 M oxyntomodulin, 0.01% (w/v) LMNG and 0.0006% (w/v) CHS before bound material was eluted in buffer containing 5 mM EGTA and 0.1 mg/mL FLAG peptide