https://bsi.inscog.eu/ 2026-06-01T23:06:57+00:00 https://bsi.inscog.eu/ https://bsi.inscog.eu/lib/exe/fetch.php?media=wiki:dokuwiki.svg text/html 2023-11-01T20:19:02+00:00 Anonymous (anonymous@undisclosed.example.com) https://bsi.inscog.eu/doku.php?id=purification:purification_carm-130-487&rev=1698869942&do=diff Purification mCARM1 construction 130-487 Sur 1l baculo. Sonication dans 50ml 50mM TrisHCl pH8, 250mM NaCl, 5% glycerol, 5mM tcep, 0.01% NP40 + anti-protéases. Centrifugation 30mn 18000rpm. Incubation EB + 2ml billes GSH-sépharose o/n 4°C. Lavages billes 2x15ml tampon. text/html 2023-11-01T20:19:02+00:00 Anonymous (anonymous@undisclosed.example.com) https://bsi.inscog.eu/doku.php?id=purification:purification_of_smprmt3_184-553&rev=1698869942&do=diff Expression & purification of 6his-SmPRMT3(1-553) Construct MGSSHHHHHHSSGTGSGENLYFQGHMSPCDQSSNYDLDMEPNDSSYFGSYGHFEIHGEMINDRVRTESYVNFILSNAEKYFKHKIILDVGSGSGILSIIAAQAGASHVYGVEAADEIYAASHETLRVNNLLERVTFIHGQAESVELPVKKVDVIISEWMGYFLFFESMLDSVLKMASKYLSRDGHIFPRHYTLNLLGVQCSEQLRKRRLEHWNNVYGYNMPALRRAALSEAHVLNLTNEHVTPPISPITILTQSFELVALDLDDMHRNRIYNLSNHCSLLCEQKFHLTIQPTTDINNNSSSSSYELDAIVGYFDVRFDDDADCKVEFSTSPTTPLTHWKQTLLFLDKPIRVKPGDKISGIITIRRATTDNRGLEINLLIGETENSLEIKQTFDLIG text/html 2023-11-01T20:19:02+00:00 Anonymous (anonymous@undisclosed.example.com) https://bsi.inscog.eu/doku.php?id=purification:storage&rev=1698869942&do=diff Flash-freezing for long-term storage Place 10 to 25 μL aliquots of the protein sample in its final buffer in PCR tubes. Submerge the tubes under the surface of liquid nitrogen using a pair of forceps until completely frozen and the transfer the tubes to a 193 K freezer for long-term storage. text/html 2023-11-01T20:19:02+00:00 Anonymous (anonymous@undisclosed.example.com) https://bsi.inscog.eu/doku.php?id=purification:tev&rev=1698869942&do=diff In-house TEV production ---------- Clones pRK792 plasmid - His-TEV(S219V)-Arg - Waugh Sequence HHHHHHHVRRRRR His-TEV(L35V/S135G/S219V)-Arg - Cabrita His-TEV plasmid - Birck pBS2440 optimized TEV plamsid - Seraphin ---------- Protocols Waugh protocol Cabrita protocol Birck protocol Seraphin protocol TEV FAQ