Table 1.

Summary of required and optional 2D-NMR experiments. See Tables S5 – S9 for detailed experimental parameters.

Experiment Code	Number of Spectra	Type of Experiment	Sample	Sampling Type	Comments
Required Experiments
D1A	30	1H,15N gHSQC1	SSS2	US3	Field dependent acquisition:20 ms in t1
D2A	39	1H,13C gHSQC	SSS	US	Field independent acquisition:128 total points in t1
D2B	32	1H,13C gHSQC	SSS	NUS4	50% NUS of D2A
D2C	41	1H,13C gHSQC	SSS	US	Field dependent acquisition:25 ms in t1
D2D	31	1H,13C gHSQC	SSS	NUS	50% NUS of D2C
D2E	26	1H,13C gHSQC	SSS	NUS	Twice the scans per increment, 50% NUS of D2C
D3A	47	1H,13C gHSQC	NIST-Fab	US	Field dependent acquisition:25 ms in t1
D3B	36	1H,13C gHSQC	NIST-Fab	NUS	50% NUS of D3A
Optional Experiments
E1	7	1H,13C gHSQC	NIST-Fab	Either	One parameter change from required experiments
E1A	45	1H,13C gHSQC	Either	NUS	Customized sampling schedule generated by individual laboratory
E1B	48	1H,13C gHSQC	Either	Either	Different temperatures:15 °C, 25 °C, 45 °C, or 50 °C
E1C	23	1H,13C sfHMQC5	Either	Either	sfHMQC pulse sequence
E2	11	1H,15N gHSQC	Either	US	One parameter change from D1A
E2N	2	1H,15N gHSQC	SSS	NUS	Customized NUS schedule generated by individual laboratory
E2A	14	1H,15N gHSQC	SSS	US	Different temperatures:15 °C, 25 °C, 45 °C, or 50 °C
E2B	8	1H,15N pHSQC	Either	US	Phase sensitive HSQCpulse sequence
E2C	11	1H,15N sfHMQC	Either	Either	SOFAST-HMQC pulse sequence

¹gHSQC = gradient selected heteronuclear single quantum coherence spectroscopy

²SSS = system suitability sample

³US = uniform sampling

⁴NUS = non-uniform sampling

⁵sfHMQC = selective optimized flip angle short transient (SOFAST) heteronuclear multiple quantum coherence spectroscopy (HMQC)