The mass of purified intact stable isotopically labeled Qconcat A protein was analyzed by MALDI (Extended Data Fig. 2E ) on a JEOL JMS-S3000 SpiralTOF mass spectrometer using the ultra-thin-layer sample preparation method60 (link),61 (link) in which α-cyano-4-hydroxycinnamic acid (Sigma) was used as the matrix. The mass of Qconcat A was internally calibrated with horse myoglobin ([m+H]+ = 16,952.5 Da). Mass calibration and background subtraction were carried out with the JEOL msTornado control software, while additional analyses were carried out with the MoverZ software62 (link). The Qconcat A protein was also characterized by peptide mapping, wherein tryptic peptides from in-gel digestion were loaded onto a PicoFrit® column (New Objective, Woburn, MA) with an integrated emitter tip (360 mm O.D., 50 mm I.D., 10 mm tip) self-packed with 6 cm of reverse-phase C18 material (ReproSil-Pur C18-AQ, 3 mm beads from Dr. Maisch GmbH), and analyzed with a LTQ Orbitrap Velos mass spectrometer (Thermo Fisher Scientific), with a Agilent 1200 series HPLC system (Agilent) and a homebuilt micro electrospray source. The purified Qconcat B was characterized by peptide mapping on a Thermo Orbitrap Fusion mass spectrometer, with a Thermo Easy-nLC 1000 HPLC and a Thermo Easy-Spray electrospray source.
Jms s3000 spiraltof mass spectrometer
Manufactured by JEOL
Sourced in Japan
The JMS-S3000 SpiralTOF mass spectrometer is a high-performance analytical instrument designed for accurate mass measurement and structural analysis of a wide range of samples. It utilizes a spiral time-of-flight (SpiralTOF) configuration to provide high mass resolution and sensitivity. The JMS-S3000 is capable of performing precise mass measurements and delivering detailed structural information for various applications.
Automatically generated - may contain errors
Lab products found in correlation
α cyano 4 hydroxycinnamic acid, by Merck Group (2 mentions)
Easy nlc 1000 hplc, by Thermo Fisher Scientific (2 mentions)
Mstornado control software, by JEOL (2 mentions)
Orbitrap fusion mass spectrometer, by Thermo Fisher Scientific (2 mentions)
Agilent 1200 series hplc system, by Agilent Technologies (2 mentions)
Ltq orbitrap velos mass spectrometer, by Thermo Fisher Scientific (2 mentions)
Picofrit column, by New Objective (2 mentions)
Mstornado control analysis, by JEOL (1 mentions)
4 protocols using jms s3000 spiraltof mass spectrometer
1
Characterization of Qconcat Proteins
2
MALDI-TOF Mass Spectrometry Protocol
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MALDI mass spectra were recorded using a JMS‐S3000 SpiralTOF™ mass spectrometer (JEOL, Tokyo, Japan). A Nd:YLF laser irradiated the spots made from the deposition of 1 μL of a matrix/salt/sample solution (5 μL of DCTB at 20 mg mL–1 in THF, 1 μL of NaTFA at 2 mg mL–1 in THF and 1 μL of the aliquot from a SEC single elution) on a target plate and allowed to air‐dry. DCTB has been expressly chosen as it provides the best signal‐to‐noise (S/N) ratios at a low laser fluence, then allowing high resolution to be achieved. In addition, it has been found that the use of alternative matrices such as dithranol or 2,5‐DHB not only dramatically deteriorates the S/N ratios but also favors the detection of the VA‐rich oligomers detrimentally to the E‐rich species, leading to overestimations of the VA content (data not shown). The so‐generated ions were accelerated by a 20 kV high voltage and went through the SpiralTOF analyzer along a spiral trajectory (approximate path length: 17 m) before their detection.32 The delay time was set according to the mass range of the sample (230 ns for fractions #3, 240 ns for fractions #2 and 350 ns for fractions #1) to keep the peak width ΔM<0.03 Da at FWHM over the mass range of interest. Calibration was performed externally using the sodium adducts of a poly(methyl methacrylate) standard (DCTB, no salt added).
3
Characterization of Qconcat Proteins
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The mass of purified intact stable isotopically labeled Qconcat A protein was analyzed by MALDI (Extended Data Fig. 2E ) on a JEOL JMS-S3000 SpiralTOF mass spectrometer using the ultra-thin-layer sample preparation method60 (link),61 (link) in which α-cyano-4-hydroxycinnamic acid (Sigma) was used as the matrix. The mass of Qconcat A was internally calibrated with horse myoglobin ([m+H]+ = 16,952.5 Da). Mass calibration and background subtraction were carried out with the JEOL msTornado control software, while additional analyses were carried out with the MoverZ software62 (link). The Qconcat A protein was also characterized by peptide mapping, wherein tryptic peptides from in-gel digestion were loaded onto a PicoFrit® column (New Objective, Woburn, MA) with an integrated emitter tip (360 mm O.D., 50 mm I.D., 10 mm tip) self-packed with 6 cm of reverse-phase C18 material (ReproSil-Pur C18-AQ, 3 mm beads from Dr. Maisch GmbH), and analyzed with a LTQ Orbitrap Velos mass spectrometer (Thermo Fisher Scientific), with a Agilent 1200 series HPLC system (Agilent) and a homebuilt micro electrospray source. The purified Qconcat B was characterized by peptide mapping on a Thermo Orbitrap Fusion mass spectrometer, with a Thermo Easy-nLC 1000 HPLC and a Thermo Easy-Spray electrospray source.
4
MALDI-TOF Mass Spectrometry of Polymer Samples
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The polymer sample was dissolved in tetrahydrofuran (∼1 mg mL−1). 5 μL of the sample solution were pipetted and mixed with 15 μL of a solution of DCTB in tetrahydrofuran at 15 mg mL−1. Five aliquots of 1 μL of the mixed solution were then deposited on a non-hydrophobic surface (384 circles) from Hudson Surface Technology (HST Inc., Old Trappan, NJ) and let to air dry. MALDI mass spectra were recorded using a JMS-S3000 SpiralTOF mass spectrometer (JEOL, Tokyo, Japan) equipped with a Nd:YLF (Neodymium-doped yttrium lithium fluoride) laser irradiating the deposits. The so-generated ions were accelerated by a 20 kV high voltage and went through the spiralTOF analyzer along a spiral trajectory (approximate path length: 17 m) before their detection.4 (link)) The delay time was set at 300 ns to keep the peak width ΔM<0.03 Da at FWHM over the mass range of interest. Calibration was performed externally and internally using the sodium adducts of poly(methyl methacrylate) 1310 g mol−1 and 2680 g mol−1 standards. MSTornado control/analysis (JEOL) was used for data acquisition while mMass 5.5.0.0 was used for data processing and artworks.25 (link))
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