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Bis[sulfosuccinimidyl] suberate (BS3) crosslinking

AS Angita Shrestha
JS Jean-Remy Sadeyen
DL Deimante Lukosaityte
PC Pengxiang Chang
AS Adrian Smith
MH Marielle Van Hulten
MI Munir Iqbal
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To determine the oligomeric structure of the recombinant H9HA containing trimerisation foldon domain, cross-linking was performed using BS3 (Thermofisher Scientific) as described by Weldon et al.45 with the following modifications. Briefly, 15 µg recombinant protein was incubated at room temperature in the presence of BS3 (final concentration 10 mM) for 1 h. Cross-linking was stopped by the addition of 1 M Tris-HCl pH 8.0 to a final concentration of 50 mM. The cross-linked products were separated on SDS gel under reducing conditions, blotted, and immunodetected using anti-H9HA monoclonal antibody44.

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