Methyl Isotope-Labeling of Proteins from NMR-Bio

Methyl groups are ideal probes for studying protein structure and function by NMR. Choose among user-friendly kits for isotope labeling of a single type or multiple types of methyl groups.

 

1. Kits for Methyl Group Assignments

Detection of long range nOesOptimized labeling of Ile, Leu, and Val for assignment. Protein was produced in U-[2H,13C,15N] M9 media supplemented with TLAM-Iγ2LVproR-U-[13C] kit.1,2

Assign specific amino acid methyl groups separately or simultaneously using kits that connect region- or stereo- specific labeled 13CH3 groups to the 15N/13C backbone nuclei via a linear 13C spin system.

 

Description  
13CH3 groups labeled      Product number
SLAM-Aβ-13C3 Alaβ 906492
SLAM-Iδ1-13C5 Ileδ1 906409
SLAM-VproR-13C4
ValproR 906417
DLAM-LVproR-13C4 LeuproR ValproR 906425
TLAM-Iδ1LVproR-U-[13C] Ileδ1 LeuproR VproR 906433
QLAM-AβIδ1LVproR-U-[13C] Alaβ Ileγ2 LeuproR ValproR 906441

 

2. Kits for Solid-State NMR studies

Achieve region and stereospecific labeling of Ile, Leu, Val, and Ala with 13CHD2 isotopomers. Acquire
high resolution solution and solid-state 1H, 13C spectra to investigate precise dynamics.

 

Description    13CHD2 groups labeled      Product number
SLAM-VproS-13CHD2 ValproS 906468
SLAM-Aβ-13CHD2 Alaβ 906476
DLAM-LVproS -13CHD2 LeuproS ValproS 906484

 

3. Kits for Large Protein Labeling

Kits for large protein labeling

Investigate protein assemblies with molecular weight up to 1 MDa.

a. Label one type of methyl group or combine kits to label multiple types.

Use a SLAM kit to achieve high incorporation of 13CH3 isotopomer on a single type of methyl group. Use in perdeuterated minimal media without detectable scrambling.

Reduce spectral overlap using combinations of single labeling (SLAM) kits and simultaneously label different types of methyl probes that are well separated in Methyl-TROSY spectra.

 

Description 
13CHD2 groups labeled      Product number
SLAM-Aβ-13CH3 Alaβ 906514
SLAM-Iγ2-13CH3 Ileγ2 906522
SLAM-VproS-13CH3 ValproS 906530
SLAM-VproR-13CH3 ValproR 906549

 

b. Detect long-range nOes or filter inter-molecular NOES in supramolecular assemblies.

Extract precise and long-range nOe distant restraints between methyl probes in perdeuterated proteins. Or, filter intra- and inter- molecular nOes and get more precise structural information to refine your homo-oligomeric protein structure.

 

Description 
13CHD2 groups labeled      Product number
DLAM-LVproS-13CH3 LeuproS ValproS 906557
DLAM-LVproR-13CH3 LeuproS ValproS 906565
DLAM-Iδ1VproS-13CH3 Ileδ1 ValproR 906573
DLAM-AβVproS-13CH3 Alaβ ValproS 906581
DLAM-Iδ1Tg-13CH3 Ileδ1 Thrg 906603
DLAM-MεVproS-13CH3 Metε ValproS 906611

 

4. Combinational Labeling Kits for Long-Range nOes Detection

Detection of long range nOesDetection of long-range nOes between methyl probes distant by up to 10 Å in large proteins and complexes.1,3

User-friendly solutions for the simultaneous isotope labeling of combinations of Ala, Ile, Leu, Met, Thr, and Val methyl groups

NMR-Bio kits include region-specific and/or stereo-specific labeling of Ile, Leu, and Val residues. Kits are provided with precise protocols extensively tested in-vivo to ensure optimal incorporation in targeted methyl groups without detectable scrambling in other positions.

The kits are specifically deuterated and supplied frozen and ready to use. All kits are calibrated for addition into deuterated M9 culture medium prior to induction. These kits incorporate 13CH3 or 13CHD­2 isotopomers into selected protein methyl groups. Protocols are provided with each kit, and each kit is designed for 1L of culture.

 

Description 
13CHD2 groups labeled      Product number
TLAM- Iδ1LVproS-13CH3 Ileδ1 LeuproS ValproS 906638
TLAM- Iδ1MεTγ-13CH3 Ileδ1 Metε Thrγ 906646
QLAM- AβIδ1LVproS-13CH3 Alaβ Ileδ1 LeuproS ValproS 906654
QLAM- Iδ1TγLVproS-13CH3 Ileδ1 Thrγ LeuproS ValproS 906662
PLAM- AβIδ1LVproSTg-13CH3 Alaβ Ileδ1 LeuproS ValproS 906670
HLAM- AβIδ1MεLVproSTg-13CH3 Alaβ Ileδ1 Metε LeuproS ValproS Thrg 906506