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[Question from NMRWiki Q&A forum] Weak NOE signals, serious t1 noise

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NMR processing:

MDD

NMR assignment:

Backbone:

Side-chains:

NOEs:

Structure from NMR restraints:

Ab initio:

Fragment-based:

Rosetta-NMR (Robetta)

Template-based:

GeNMR

I-TASSER

Refinement:

Amber

Structure from chemical shifts:

Fragment-based:

Homology-based:

Torsion angles from chemical shifts:

Preditor

TALOS

Promega- Proline

Secondary structure from chemical shifts:

CSI (via RCI server)

TALOS

MICS caps, β-turns

d2D

PECAN

Flexibility from chemical shifts:

RCI

Interactions from chemical shifts:

HADDOCK

Chemical shifts re-referencing:

NMR model quality:

NOEs, other restraints:

Chemical shifts:

RDCs:

Pseudocontact shifts:

Protein geomtery:

SAVES2 or SAVES4

Quality Control Check

NMR spectrum prediction:

FANDAS

MestReS

V-NMR

Flexibility from structure:

Backbone S2

Methyl S2

B-factor

Molecular dynamics:

Gromacs

Amber

Antechamber

Chemical shifts prediction:

From structure:

Shiftx2

Sparta+

Camshift

CH3shift- Methyl

ArShift- Aromatic

ShiftS

Proshift

PPM

CheShift-2- Cα

From sequence:

Shifty

Camcoil

Poulsen_rc_CS

Disordered proteins:

MAXOCC

Format conversion & validation:

CCPN

From NMR-STAR 3.1

Validate NMR-STAR 3.1

NMR sample preparation:

Protein disorder:

DisMeta

Protein solubility:

Isotope labeling:

Solid-state NMR:

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10-26-2014, 12:20 AM

nmrlearner

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Weak NOE signals, serious t1 noise

Weak NOE signals, serious t1 noise

Since my university does not have a professional spectroscopist, I am a new learner of NMR, and I really have no one else to ask...

I have a very concentrated sample (small molecule, MW < 300, 100 mg / mL), and I am running a 2D NOESY experiment (pulseprogram = noesyph in Bruker 400 MHz). However, the NOE signals that I am looking for are very weak, hidden by serious t1 noise (since the signal concerned is a CH3 group, very strong proton signal). Besides tricks like symmetrization, what else can I do to reduce the t1 noise or strengthen my NOE signal?

Shall I:1. Increase ns (no of scan, currently ns = 8)?2. Increase d1 (relaxation delay, currently d1 = 3 s)?3. Increase d8 (mixing time, currently d8 = 1.5 s)?

Thanks a lot for your help!

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