TRAF2 saxs H4
Description
Small Angle X-ray Scattering data of the protein TRAF2 (266-501) - HPLC mode, concentration 4 mg/ml
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Steps to reproduce
The sequence coding for the residues 266-501 (C-Ter) of TRAF2 was cloned in the pET21(A) vector. Plasmids were used to transform Escherichia coli strain C41(DE3) competent cells. Protein expression was induced by adding isopropyl--D-thiogalactropyranoside (IPTG) to a final concentration of 0.5 M. Bacteria were grown in LB medium with chloramphenicol 34 mg/ml and Ampicillin 100 mg/ml. Cells were harvested and resuspended in a buffer containing 50 mM Tris-HCL pH 8.0, 500 mM NaCl and protease inhibitors, treated with 4 g/ml DNase, 40 mM MgSO4 and 50 g/ml lysozyme and disrupted by French Press. After elimination of debris by high-speed centrifugation, TRAF2 C-TER was purified using Ni-NTA (HisTrap FF crude GE Healthcare), using 500 mM Imidazole for their elution. After a gel filtration step, proteins were stored in 50 mM BIS-TRIS pH 8.0, 200 mM NaCl, 10 mM dithiothreitol (DTT). SAXS data were recorded at BM29 ESRF in Grenoble, HPLC mode at the concentration of 4 mg/ml.
Institutions
- Istituto di Biofisica Consiglio Nazionale delle Ricerche Sede secondaria di MilanoLombardia, Milano