New study challenges our understanding of the immunesystem NatureComms
, 20 mL 10% glucose solution and 2 mL AMP . The culture was grown at 37 °C and 120 RPM to an optical density 600 ~0.6 at which point the expression was induced with 0.5 mM isopropyl β-D-1-thiogalactopyranoside at 25 °C O/N. The following day, cells were harvested and resuspended in lysis buffer supplemented with 5 mM β-mercaptoethanol prior to sonication for 5 min at 8 watts in pulses of 10 s and 10 s of rest.
8 mg/mL was conjugated with 7x excess of DBCO-S1-handle with an extension carrying P3 docking site by incubating O/N at RT and protected from light. To remove unreacted IgG as well as DNA, mIgG2a-DNA-purification was performed on an ÄKTA pure system using 1 mL RESOURCE Q column. Appropriate fractions were concentrated and buffer-exchanged into PBS via 100 kDa MWCO centrifugal filter. The mIgG2a-DNA conjugate was stored at 4 °C at a concentration of 3.3 μM.
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