The crystals were grown using the in meso approach [62 ,63 (link),64 ], similarly to our previous work [41 (link),44 (link)]. The solubilized protein in the crystallization buffer was added to the mono-oleoyl-formed lipidic phase (Nu-Chek Prep, Elysian, MN, USA). Crystallization trials were set up using the NT8 robotic system (LCP version, Formulatrix, Bedford, MA, USA). The crystals were grown at 22 °C and reached the final size of ~100 µm within 1 week. The crystals were obtained using the precipitant solution consisting of 1.2 M KH2PO4/Na2HPO4 pH 4.6. Before harvesting, the crystals were incubated for 5 min in the respective precipitant solutions supplemented with 20% glycerol. All crystals were harvested using micromounts (MicroLoops HT, MiTeGen, Ithaca, NY, USA), then flash-cooled and stored in liquid nitrogen.
Microloops ht
Manufactured by MiTeGen
Sourced in United States
MicroLoops HT are a set of pre-mounted, disposable protein crystallization sample holders designed for high-throughput screening. They provide a stable and controlled environment for protein crystal growth experiments.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using microloops ht
1
In meso protein crystal growth
2
Crystallization of Membrane Proteins
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The crystals were grown using the in meso approach [17] [18] (link)[19] , similarly to our previous work [13] (link). The solubilized protein in the crystallization buffer was added to the monooleoyl-formed lipidic phase (Nu-Chek Prep, Elysian, MN, USA). Crystallization trials were set up using the NT8 robotic system (LCP version, Formulatrix, Bedford, MA, USA). The crystals were grown at 22 • C and reached the final size of ~100 µm within 3 months. The crystals were obtained using the precipitant solution consisting of 1 M KH 2 PO 4 /Na 2 HPO 4 pH 5.2 and 5 mM NaNO 3 . Before harvesting, the crystals were incubated for 5 min in the respective precipitant solutions supplemented with 20% glycerol. All crystals were harvested using micromounts (MicroLoops HT, MiTeGen, Ithaca, NY, USA), then flash-cooled and stored in liquid nitrogen.
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