Tka x cad system

CaM (bovine brain) was obtained from Athens Research (Athens, GA, USA). Cross-linkers were purchased from Thermo Fisher Scientific (Dreieich, Germany) (s-GMBS, BS³ and photo-Met) or synthesized in house (DSBU) [42 (link)]. The proteases trypsin (porcine) and GluC were obtained from Promega GmbH (Mannheim, Germany). Chemicals were obtained from Sigma Aldrich (Taufkirchen, Germany), Merck (Darmstadt, Germany), and VWR (Darmstadt, Germany). Prestained Protein Ladder was purchased from Thermo Fisher Scientific. For HPLC, solvents were LC-MS grade (VWR, Darmstadt, Germany). MilliQ water was produced by a TKA X-CAD system (Thermo Fisher Scientific, Niederelbert, Germany).

Agonists GW0742 and GW1516 were purchased from Sigma-Aldrich (Taufkirchen, Germany). Nano-HPLC solvents were LC-MS grade (VWR, Darmstadt, Germany), water was purified with a TKA X-CAD system (Thermo Fisher Scientific, Bremen, Germany). Trypsin (cleaving C-terminally of lysine and arginine), GluC (cleaving C-terminally of glutamate and aspartate), and ProTEV Plus (cleaving C-terminally of ENLYFQ(G/S)) were obtained from Promega (Mannheim, Germany). The cross-linker BS²G-D₀/D4 was obtained from Thermo Fisher Scientific (Rockford, IL). The urea-linker was synthesized and purified in-house.[13 (link)] Bpa was obtained from Bachem (Bubendorf, Switzerland). Tryptone, yeast extract, antibiotics, TCEP, and IPTG were purchased from Roth (Karlsruhe, Germany). Iodacetamide, DTT, D-desthiobiotin, DMTMM, and all other chemicals (Sigma-Aldrich, Taufkirchen, Germany) were obtained at the highest available purity.