Purified GABA-A receptor from affinity chromatography was mixed with Fab in a 3:1 w:w ratio and injected over a Superose 6 Increase 10/300 GL column (GE Healthcare) equilibrated in SEC buffer. Peak fractions were assayed by SEC, monitoring tryptophan fluorescence. The peak fraction was concentrated ten-fold to 5-6 mg/ml. Three μL of purified GABA-A receptor + GABA + flumazenil + Fab complex was applied to glow-discharged gold R1.2/1.3 200 mesh holey carbon grids (Quantifoil) and immediately blotted for 3 s at 100% humidity/4 °C, then plunge-frozen into liquid ethane cooled by liquid nitrogen using a Vitrobot Mark IV (FEI).
Glow discharged gold r1.2 1 3 200 mesh holey carbon grids
Manufactured by Quantifoil
Glow-discharged gold R1.2/1.3 200 mesh holey carbon grids are a type of lab equipment used in electron microscopy. They provide a substrate for sample preparation, with a carbon film that contains regularly spaced holes. The grids are made of gold and have a mesh size of 200.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using glow discharged gold r1.2 1 3 200 mesh holey carbon grids
1
Cryo-EM Structure of GABA-A Receptor
2
Cryo-EM Structure of GABA-A Receptor
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