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All the molecules were purified by twice recrystallization in a yield of higher than 80% (Supplementary Fig. 1 53.
MHC molecules were purified by affinity chromatography using the anti-IAb monoclonal antibody Y3JP.
The Mamu-DR/P65 molecules were purified via two-round procedures as we recently described [22].
The excess biotin was removed by overnight PBS dialysis, the biotinylated molecules were purified by passing through an avidin affinity column (Pierce) as following to the supplier's recommendations.
Radiolabelled RNA11 molecules were purified using RNeasy spin columns (QIAGEN).
Sulfated molecules were purified using Sephadex G10 size exclusion chromatography.
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In the next step, the cross-linked RNA molecules are purified via immunoprecipitation of the RBP and sequenced.
The poly(A) containing mRNA molecules was purified using Sera-mag Magnetic Oligo dT) Beads (Illumina) from 20 mg total RNA of each sample.
Briefly, starting from 1 μg of total RNA, the poly-A containing mRNA molecules was purified using poly-T oligo-attached magnetic beads.
This modified I-Ad molecule was purified using preparative IEF and one fraction, after removal of the leucine zipper tails, produced crystals suitable for X-ray crystallographic analysis.
Material eluted from murine CD1d molecules that were purified from transfected insect cells included mainly PC and phosphatidylethanolamine (PE) [7].
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