3.2mm zirconia thin wall mas rotors bruker biospin  (Bruker Corporation)

Journal: Journal of magnetic resonance (San Diego, Calif. : 1997)

Article Title: Sensitivity Boosts by the CPMAS CryoProbe for Challenging Biological Assemblies

doi: 10.1016/j.jmr.2019.106680

Figure Lengend Snippet: a),b) 1D and 2D MAS NMR spectra of Kif5b/MT assembly acquired with the 3.2 mm CPMAS CryoProbe at 14.1 T (purple) and 3.2 mm EFree probe at 20.0 T (black): a) 13C CPMAS; b) CORD (top) and NCACX (bottom). c) Expansions of the CORD spectra showing selected regions containing the aliphatic and aromatic sidechain correlations. d) Selected 1D traces through the direct (top) and indirect (bottom) dimensions of the CORD spectrum, for chemical shifts shown as green dashed lines. The CPMAS spectra were collected with 64 and 512 scans, and the total acquisition time was 16 ms. The SNR = 135 and 136; the effective SNR per mg of protein per square root of scan and corrected for the B0 = 2.2 and 0.7 for CPMAS CryoProbe and EFree probe spectra, respectively. The CORD spectra were collected with 36 and 192 scans, the recycle delays were 2 and 3 s; the total experiment times were 1.6 and 2.5 days. The SNR (1st FID) = 60 and 59 with equivalent processing. The effective SNR per mg of protein per square root of scan and corrected for the B0 = 1.3 and 0.5 for CPMAS CryoProbe and EFree probe spectra, respectively. The NCACX spectra were collected with 256 and 3072 scans, the recycle delays were 3 and 2 s; the total experiment times were 17 and 139 hours. The SNR (1st FID) = 13 for both data sets. The effective SNR per mg of protein per square root of scan and corrected for the B0 = 0.1 and 0.02 for CPMAS CryoProbe and EFree probe spectra, respectively. The MAS frequency in both cases was 14 kHz. The estimated amount.of U-13C,15N-Kif5b in the samples for experiments with the CPMAS CryoProbe and EFree probe was 7.5 and 5.0 mg; the total amount of assemblies packed in the rotors was 82.4 and 55.0 mg.

Article Snippet: The resulting complex was pelleted by ultracentrifugation at 156,800 g (60,000 rpm) at 20 °C for 1 hour; the hydrated pellet (82.4 mg total weight, corresponding to ~7.5 mg of U- 13 C, 15 N-enriched Kif5b) was transferred into a thin-wall 3.2 mm Bruker CPMAS CryoProbe rotor.

Techniques:

Journal: Journal of magnetic resonance (San Diego, Calif. : 1997)

Article Title: Sensitivity Boosts by the CPMAS CryoProbe for Challenging Biological Assemblies

doi: 10.1016/j.jmr.2019.106680

Figure Lengend Snippet: 1D and 2D MAS NMR spectra of conical assembly of U-13C,15N-CA (estimated protein amount: 65 mg) acquired with the 3.2 mm CPMAS CryoProbe: a),b) 15N and 13C CPMAS; c) CORD (top) and NCACX (bottom). d) Expansions of the CORD spectrum showing the Ser-Thr region (top) and the aromatic-to-aromatic sidechain correlations (bottom). The CPMAS spectra were collected with 4 and 8 scans, and the total acquisition time was 21 and 16 ms, respectively. The SNR = 26 and 237; the effective SNR per mg of protein per square root of scan = 0.2 and 1.0, respectively. The CORD spectrum was collected with 2 scans, the acquisition time in the direct dimension was 18 ms; the total experiment time was 1.7 hours. The SNR (1st FID) = 114.7. The NCACX spectrum was collected with 40 scans, the acquisition time in the direct dimension was 17 ms; the total experiment time was 2.1 hours. The MAS frequency was 14 kHz. The SNR (1st FID) = 26. The MAS frequency was 14 kHz.

Article Snippet: The resulting complex was pelleted by ultracentrifugation at 156,800 g (60,000 rpm) at 20 °C for 1 hour; the hydrated pellet (82.4 mg total weight, corresponding to ~7.5 mg of U- 13 C, 15 N-enriched Kif5b) was transferred into a thin-wall 3.2 mm Bruker CPMAS CryoProbe rotor.

Techniques:

Journal: Journal of magnetic resonance (San Diego, Calif. : 1997)

Article Title: Sensitivity Boosts by the CPMAS CryoProbe for Challenging Biological Assemblies

doi: 10.1016/j.jmr.2019.106680

Figure Lengend Snippet: Selected 2D planes for the 3D NCACX (purple) and NCOCX (black) spectra of Kif5b/MT assembly acquired with the 3.2 mm CPMAS CryoProbe at 14.1 T. The NCACX spectrum was acquired with 48 scans per FID, as 1818×64×50 (t3×t2×t1) complex matrix. The recycle delay was 2.7 s, and the total experiment time was 4.8 days. The effective SNR per mg of protein per square root of scan = 0.14. The NCOCX spectrum was acquired with 96 scans per FID, as 1818×36×50 (t3×t2×t1) complex matrix. The recycle delay was 3.0 s, and the total experiment time was 6 days. The effective SNR per mg of protein per square root of scan = 0.23.

Article Snippet: The resulting complex was pelleted by ultracentrifugation at 156,800 g (60,000 rpm) at 20 °C for 1 hour; the hydrated pellet (82.4 mg total weight, corresponding to ~7.5 mg of U- 13 C, 15 N-enriched Kif5b) was transferred into a thin-wall 3.2 mm Bruker CPMAS CryoProbe rotor.

Techniques:

Journal: Journal of magnetic resonance (San Diego, Calif. : 1997)

Article Title: Sensitivity Boosts by the CPMAS CryoProbe for Challenging Biological Assemblies

doi: 10.1016/j.jmr.2019.106680

Figure Lengend Snippet: 1D and 2D MAS NMR spectra of tubular assembly of U-15N-CA (estimated protein amount: 65 mg) acquired with the 3.2 mm CPMAS CryoProbe: a) 15N CPMAS; b) 1H-15N DANTE-HETCOR; c) 15N-15N PDSD; d) NCO (left) and NCA (right). The 15N CPMAS spectrum was collected with 4 scans, the total acquisition time was 33 ms. The SNR = 59; the effective SNR per mg of protein per square root of scan = 1.3. Expansions around selected regions of the spectrum illustrate the high spectral resolution. The 1H-15N DANTE-HETCOR spectrum was collected with 48 scans; the total experiment time was 5.2 hours. The SNR (1st FID) = 20. The 15N-15N PDSD spectrum was collected with 64 scans; the total experiment time was 13.75 hours. The SNR (1st FID) - 128. The spectrum was processed with 90- and 60-degree shifted sinebell functions (gray and purple contours, respectively). The NCO and NCA spectra were both acquired with 512 scans, as 2048×54 and 2048×62 (t2×t1) complex matrices; and the total experiment time was 23.5 and 27 hours, respectively. The SNR (1st FID) was 5 (NCA) and 7.6 (NCO).

Article Snippet: The resulting complex was pelleted by ultracentrifugation at 156,800 g (60,000 rpm) at 20 °C for 1 hour; the hydrated pellet (82.4 mg total weight, corresponding to ~7.5 mg of U- 13 C, 15 N-enriched Kif5b) was transferred into a thin-wall 3.2 mm Bruker CPMAS CryoProbe rotor.

Techniques:

Journal: Journal of magnetic resonance (San Diego, Calif. : 1997)

Article Title: Sensitivity Boosts by the CPMAS CryoProbe for Challenging Biological Assemblies

doi: 10.1016/j.jmr.2019.106680

Figure Lengend Snippet: 2D and 3D MAS NMR spectra of U-13C,15N huPrP23–144 fibrils recorded using the 3.2 mm CPMAS CryoProbe at 600 MHz and 13 kHz MAS rate. (a) 2D NCA spectrum recorded as a 1024×47 (t2×t1) complex matrix with acquisition times of 15.3 ms (t1) and 22.5 ms (t2), 16 scans per row, 2.0 s recycle delay and a total measurement time of 50 min. (b) A reference 2D NCA spectrum of U-13C,15N huPrP23–144 fibrils recorded with a total measurement time of 7 h using the 3.2 mm BioMAS probe at 500 MHz and 11.111 kHz MAS rate. The spectrum was collected with the same recycle delay and acquisition times in t1 and t2 as the 2D NCA spectrum in panel (a) and processed in identical manner. (c) 2D N(CA)CX spectrum recorded with a DARR 13C-13C mixing time of 10 ms as a 681×57 (t2×t1) complex matrix with acquisition times of 18.7 ms (t1) and 15.0 ms (t2), 16 scans per row, 3.5 s recycle delay and a total measurement time of 1.8 h. (d) 2D N(CO)CX spectrum recorded with a DARR 13C-13C mixing time of 50 ms as a 681×32 (t2×t1) complex matrix with acquisition times of 12.3 ms (t1) and 15.0 ms (t2), 64 scans per row, 3.5 s recycle delay and a total measurement time of 4 h. (e) Representative planes corresponding to 15N frequencies of residues A115, A116 and A117 from a 3D NCOCX spectrum recorded with a DARR 13C-13C mixing time of 50 ms as a 681×64×10 (t3×t2×t1) complex matrix with acquisition times of 4.6 ms (t1), 12.3 ms (t2) and 15.0 ms (t3), 16 scans per row, 3.5 s recycle delay and a total measurement time of 40 h. (f) Representative planes corresponding to 15N frequencies of residues A115, A116 and A117 from a 3D NCACX spectrum recorded with a DARR 13C-13C mixing time of 50 ms as a 388×40×16 (t3×t2×t1) complex matrix with acquisition times of 6.2 ms (t1), 6.2 ms (t2) and 15.0 ms (t3), 16 scans per row, 3.5 s recycle delay and a total measurement time of 40 h.

Article Snippet: The resulting complex was pelleted by ultracentrifugation at 156,800 g (60,000 rpm) at 20 °C for 1 hour; the hydrated pellet (82.4 mg total weight, corresponding to ~7.5 mg of U- 13 C, 15 N-enriched Kif5b) was transferred into a thin-wall 3.2 mm Bruker CPMAS CryoProbe rotor.

Techniques:

Journal: Journal of magnetic resonance (San Diego, Calif. : 1997)

Article Title: Sensitivity Boosts by the CPMAS CryoProbe for Challenging Biological Assemblies

doi: 10.1016/j.jmr.2019.106680

Figure Lengend Snippet: a) 1D MAS CP spectra of HET-s (218–289) fibrils acquired with a single scan at 14.1 T with the 3.2 mm CPMAS CryoProbe (blue) and 3.2 mm EFree probe (black). (b-c) 2D PDSD spectra of HET-s (218–289) fibrils recorded at 14.1 T with the 3.2 mm CPMAS CryoProbe (b) and 3.2 mm EFree probe (c). The CC spectra were collected with 1 scan for the BioSolids CryoProb™e and 16 scans for the EFree probe, The recycle delays were respectively 3.5 and 2 s; the total experiment times were 1.3 and 10 hours. The MAS frequency in both cases was 16 kHz. The estimated amount of HET-s (218–289) fibrils in the samples for experiments with the CPMAS CryoProbe and EFree probe was 21.4 and 10.0 mg.

Article Snippet: The resulting complex was pelleted by ultracentrifugation at 156,800 g (60,000 rpm) at 20 °C for 1 hour; the hydrated pellet (82.4 mg total weight, corresponding to ~7.5 mg of U- 13 C, 15 N-enriched Kif5b) was transferred into a thin-wall 3.2 mm Bruker CPMAS CryoProbe rotor.

Techniques: