Binding studies were performed at 298K in buffer (50 mM Tris-d11, pH 7.5, 10%v/v D2O, 10 mM CaCl2) using CPMG NMR spectroscopy (56 ) and a water suppression pulse sequence. Concentrated R132H IDH1 (1.4 mM stock) was titrated (in 2.86 μL increments) into a mixture (160 μL total volume) containing a 2OG derivative (50 μM) in 3 mm diameter MATCH NMR tubes (Bruker).
Match nmr tubes
MATCH NMR tubes are specialized laboratory equipment designed for nuclear magnetic resonance (NMR) spectroscopy. They provide a precise and stable environment for sample analysis. The tubes are manufactured to high standards to ensure consistent performance and reliable results.
2 protocols using match nmr tubes
NMR Characterization of R132H IDH1
Binding studies were performed at 298K in buffer (50 mM Tris-d11, pH 7.5, 10%v/v D2O, 10 mM CaCl2) using CPMG NMR spectroscopy (56 ) and a water suppression pulse sequence. Concentrated R132H IDH1 (1.4 mM stock) was titrated (in 2.86 μL increments) into a mixture (160 μL total volume) containing a 2OG derivative (50 μM) in 3 mm diameter MATCH NMR tubes (Bruker).
NMR-based Kinetic Assay for 2HG Oxidation
The CPMG (Carr–Purcell–Meiboom–Gill) displacement experiments were conducted using the AV700 instrument and 3 mm MATCH NMR tubes32 (link). The PROJECT-CPMG sequence (90°x–[τ–180°y–τ–90°y–τ–180°y–τ]n–acq) was applied as described by Aguilar et al.56 Typical experimental parameters were as follows: total echo time 48 ms (τ=2 ms, n=6), acquisition time 2.94 s, relaxation delay 2 s and number of transients 128. Water suppression was achieved by presaturation.
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