Peter van Mourik

78 Chapter 4 Table 1: overview of the stock preparation of the individual medium components with all relevant ordering and dissolvent information: Compound Amount Concen-tration stock End concen-tration colon medium +/+ Storage Ad-DF+++ Add 5 mL Glutamax (100 x) 40% 8-12 weeks at 4 °C Add 5 mL HEPES (1M) Add 5 mL Pen/Strep (10,000 U/mL / 100 x) B27 10 mL ♢ direct use from bottle - 2% 1 yr at -20 °C N-acetylcysteine 815 mg Nac in 10 mL H 2 0 500 mM 1.25 mM 1 yr at -20 °C hEGF Add 2 mL filter sterilized PBS0-BSA (0,1 %) to 1 vial hEGF 0.5 mg/mL 10.000 x 50 ng/mL 1 yr at -20 °C Nicotinamide 6 gr nicotinamide in 50 mL PBS0 1 M 100 x 10 mM 4 months at -20 °C A83-01 10 mg A83-01 in 5 mL DMSO 5 mM 10.000 x 500 nM 3 months at -20 °C P38 inhibitor (SB202190) Add 500 µL DMSO to one bottle of 5 mg P38-inhibitor 30 mM 3000 x 3 months at -20 °C Gastrin Mix 0.5 mg gastrin to 2.383 mL PBS0 100 µM 5 nM 3 months at -20 °C R-spo3 Reconstitute in PBS0 Varies per batch 250 ng/ mL 3 months at -80 °C WCM Collect from producing cell-line (see 2.1) 2 x 50% 2 months at 4 °C NCM Collect from producing cell-=line (see 2.2) 10 x 10% 1 yr at 20 °C