600 and 700 mhz spectrometers, by Bruker - Product details

1

NMR Characterization of Guanine Residues

Check if the same lab product or an alternative is used in the 5 most similar protocols

NMR experiments were performed on Bruker 600 and 700 MHz spectrometers at 25°C, unless otherwise specified. Resonances for guanine residues were assigned unambiguously by using site-specific low-enrichment ¹⁵N-labelling (54 (link)) and site-specific ²H labelling (55 (link)). Spectral assignments were assisted by NOESY, COSY, TOCSY and ¹³C-¹H-HSQC, as previously described (56 (link),57 (link)). All spectral analyses were performed using the program FELIX (Felix NMR, Inc.).

Lim K.W., Jenjaroenpun P., Low Z.J., Khong Z.J., Ng Y.S., Kuznetsov V.A, & Phan A.T. (2015). Duplex stem-loop-containing quadruplex motifs in the human genome: a combined genomic and structural study. Nucleic Acids Research, 43(11), 5630-5646.

+ Open protocol

+ Expand

2

NMR Characterization of Guanine Residues

Check if the same lab product or an alternative is used in the 5 most similar protocols

Nuclear magnetic resonance (NMR) experiments were performed on Bruker 600 and 700 MHz spectrometers at 25°C, unless otherwise specified. Resonances for guanine residues were assigned unambiguously by using site-specific low-enrichment 15N-labelling (61 (link)) and site-specific 2H labeling (62 (link)). Spectral assignments were assisted by NOESY, COSY, TOCSY and 13C-1HHSQC, as previously described (63 (link),64 (link)). All spectral analyses were performed using the program FELIX (Felix NMR, Inc.).

Papp C., Mukundan V.T., Jenjaroenpun P., Winnerdy F.R., Ow G.S., Phan A.T, & Kuznetsov V.A. (2023). Stable bulged G-quadruplexes in the human genome: identification, experimental validation and functionalization. Nucleic Acids Research, 51(9), 4148-4177.

+ Open protocol

+ Expand

3

Phosphorylation and Calorimetric Analysis of CrkII

Check if the same lab product or an alternative is used in the 5 most similar protocols

For NMR, all samples were dialyzed in NMR buffer 50 mM potassium phosphate (pH 6.8), 140 mM NaCl and 1 mM β-mercaptoethanol). NMR experiments were performed on Varian 800- and 600-MHz and Bruker 700- and 600-MHz spectrometers. Abl kinase domain (Abl KD) was expressed and purified as described before (41 (link), 42 (link)). Purified and Tev-processed Crk II was phosphorylated by the addition of catalytic amounts of Abl KD in buffer (50mM KPi (pH 6.8), 150mM NaCl, 1mM β-mercaptoethanol) supplemented with 5 mM ATP and 10 mM MgCl₂. The reaction was carried out at room temperature. Stoichiometry of phosphorylation at Y221 was >95% by NMR.
All calorimetric titrations were performed on an iTC200 microcalorimeter (GE). Unphosphorylated or phosphorylated CrkII was at 40 µM in the sample cell and the PPII ligand was at 400 µM in the injection syringe. Ligand was dissolved in the flow through of the last buffer exchange. Data for the preliminary injection, which are affected by diffusion of the solution from and into the injection syringe during the initial equilibration period, were discarded. The data were fitted with Origin 7.0.

Sriram G., Jankowski W., Kasikara C., Reichman C., Saleh T., Nguyen K.Q., Li J., Hornbeck P., Machida K., Liu T., Li H., Kalodimos C.G, & Birge R.B. (2014). Iterative Tyrosine Phosphorylation Controls Non-canonical Domain Utilization in Crk. Oncogene, 34(32), 4260-4269.

+ Open protocol

+ Expand

4

Phosphorylation and Calorimetric Analysis of CrkII

Check if the same lab product or an alternative is used in the 5 most similar protocols

For NMR, all samples were dialyzed in NMR buffer 50 mM potassium phosphate (pH 6.8), 140 mM NaCl and 1 mM β-mercaptoethanol). NMR experiments were performed on Varian 800- and 600-MHz and Bruker 700- and 600-MHz spectrometers. Abl kinase domain (Abl KD) was expressed and purified as described before (41 (link), 42 (link)). Purified and Tev-processed Crk II was phosphorylated by the addition of catalytic amounts of Abl KD in buffer (50mM KPi (pH 6.8), 150mM NaCl, 1mM β-mercaptoethanol) supplemented with 5 mM ATP and 10 mM MgCl₂. The reaction was carried out at room temperature. Stoichiometry of phosphorylation at Y221 was >95% by NMR.
All calorimetric titrations were performed on an iTC200 microcalorimeter (GE). Unphosphorylated or phosphorylated CrkII was at 40 µM in the sample cell and the PPII ligand was at 400 µM in the injection syringe. Ligand was dissolved in the flow through of the last buffer exchange. Data for the preliminary injection, which are affected by diffusion of the solution from and into the injection syringe during the initial equilibration period, were discarded. The data were fitted with Origin 7.0.

Sriram G., Jankowski W., Kasikara C., Reichman C., Saleh T., Nguyen K.Q., Li J., Hornbeck P., Machida K., Liu T., Li H., Kalodimos C.G, & Birge R.B. (2014). Iterative Tyrosine Phosphorylation Controls Non-canonical Domain Utilization in Crk. Oncogene, 34(32), 4260-4269.

+ Open protocol

+ Expand

600 and 700 mhz spectrometers

Lab products found in correlation

Close spelling variants of this product

4 protocols using 600 and 700 mhz spectrometers

NMR Characterization of Guanine Residues

NMR Characterization of Guanine Residues

Phosphorylation and Calorimetric Analysis of CrkII

Phosphorylation and Calorimetric Analysis of CrkII

About PubCompare

Ready to get started?