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Autoflex system

Manufactured by Bruker
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Sourced in United States, Germany

The Autoflex system is a versatile mass spectrometry platform designed for a wide range of analytical applications. The core function of the Autoflex system is to provide high-performance mass analysis and detection capabilities for various sample types.

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Lab products found in correlation

Mtp 384 ground steel maldi plate, by Bruker (2 mentions) Flexanalysis 3, by Bruker (2 mentions) Deuterated nmr solvents, by Cambridge Isotopes (1 mentions) Formic acid hcooh, by Merck Group (1 mentions) Ics 5000 hplc system, by Thermo Fisher Scientific (1 mentions) Dionex carbopac pa200 column, by Thermo Fisher Scientific (1 mentions) Carbopac pa200, by Thermo Fisher Scientific (1 mentions) Chromeleon software version 7, by Thermo Fisher Scientific (1 mentions) Axima resonance maldi quadrupole ion trap tof instrument, by Shimadzu (1 mentions)

7 protocols using autoflex system

1

Fungal Identification Using MALDI-TOF MS

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All isolated fungi were identified using MALDI-TOF MS. Isolated fungal colonies on SDA plates were identified using MALDI-TOF MS. Samples were prepared following the protocol for fungi identification as by the manufacturer. Spectra were obtained with an Autoflex system (Bruker, Leipzig, Germany). Identification was performed using the MBT software, with the latest upgrade of the Bruker fungi library (MBT 7854 Species/Entry List September 2018). In parallel, spectra were also analyzed also using the MSI library [17 (link)] accessed online; https://biological-mass-spectrometry-identification.com/msi/-LIBRARY (7 September 2022). Part of the fungi isolates were also analyzed using the MALDI-TOF-MS system in another medical center (Sheba Medical Center).
Frenkel M., Serhan H., Blum S.E., Fleker M., Sionov E., Amit S., Gazit Z., Gefen-Halevi S, & Segal E. (2022). What Is Hiding in the Israeli Mediterranean Seawater and Beach Sand. Journal of Fungi, 8(9), 950.
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2

MALDI-TOF Mass Spectrometry of Assay Supernatant

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Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry was performed using a matrix of 9 mg/ml 2,5-dihydroxybenzoic acid (DHB) (Sigma-Aldrich, P/N: 149357) dissolved in 0.2% trifluoroacetic acid (TFA). 2 μL of the DHB matrix was mixed with 2 μL assay supernatant and placed on a Bruker MTP 384 ground steel MALDI plate (Bruker Daltonics, Bremen, Germany P/N: 8280784) and dried under a stream of air prior to data collection. Experiments were performed on a Bruker Daltonics Autoflex system (Billerica, MA, U.S.A.). The data acquired over the range of m/z 0 to 3000 were collected in positive-ion mode by averaging 2000 laser shots using the lowest energy sufficient to obtain adequate signal-to-noise ratios.
Li J., Solhi L., Goddard-Borger E.D., Mathieu Y., Wakarchuk W.W., Withers S.G, & Brumer H. (2021). Four cellulose-active lytic polysaccharide monooxygenases from Cellulomonas species. Biotechnology for Biofuels, 14, 29.
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3

MALDI-TOF and MALDI-QIT-TOF Mass Spectrometry Analysis

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MS analysis was carried out using a matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS instrument with an Autoflex system from Bruker Daltonics (Bruker, Germany). Tandem mass spectrometric analysis was performed using an Axima Resonance MALDI-quadrupole ion trap (QIT)-TOF instrument (Shimadzu, UK). These mass spectrometers were operated in positive-ion reflectron mode. The matrix solution used was 10 mg/ml DHB in 50% methanol. Data acquisition and processing in MS and MS/MS analysis were performed with flexAnalysis 3.3 software (Bruker, Germany) and Launchpad 2.9.3 software (Kratos Analytical Ltd., UK), respectively. The mass spectrometric analysis was triplicated for all samples.
Park H.M., Kim Y.W., Kim K.J., Kim Y.J., Yang Y.H., Jin J.M., Kim Y.H., Kim B.G., Shim H, & Kim Y.G. (2014). Comparative N-Linked Glycan Analysis of Wild-Type and α1,3-Galactosyltransferase Gene Knock-Out Pig Fibroblasts Using Mass Spectrometry Approaches. Molecules and Cells, 38(1), 65-74.
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4

Inert Atmosphere Synthesis and Characterization

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All air- and water-sensitive procedures were carried out either in a Vacuum Atmospheres glovebox under nitrogen (0.5−10 ppm of O2 for all manipulations) or using standard Schlenk techniques under nitrogen. Deuterated NMR solvents were purchased from Cambridge Isotopes Laboratories. Benzene and dichloromethane-d2 were dried over sodium benzophenone ketyl and distilled prior to use. Formic acid (HCOOH) was purchased from Sigma-Aldrich and used under a N2 atmosphere without further purifications. Acetic acid was dried over CaH2 and distilled prior to use. The integrity of this material was checked regularly by NMR. Cyclooctadiene iridium chloride dimer (7) was purchased from Sigma-Aldrich and used as received. Sonication procedures were done in a VWR desktop sonic cleaner bath. Silver trifluoromethylsulfonate was purchased from Alfa Aesar and used as received. 1H, 13C, and 31P NMR spectra were obtained on Varian 600 or 500 MHz spectrometers with chemical shifts reported in units of ppm. All 1H chemical shifts were referenced to the residual 1H solvent (relative to TMS). NMR spectra were taken in 8-inch J-Young tubes (Wilmad) with Teflon valve plugs. All spectra were processed using MesRe Nova (v.11.0.4–11998). MALDI mass spectrometry was conducted on a Bruker Autoflex system.
Lauridsen P.J., Lu Z., Celaje J.J., Kedzie E.A, & Williams T.J. (2018). Conformational Twisting of a Formate-Bridged Diiridium Complex Enables Catalytic Formic Acid Dehydrogenation. Dalton transactions (Cambridge, England : 2003), 47(38), 13559-13564.
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5

Structural Analysis of Mixed-Linkage Glucans

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High performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) was performed using a Dionex ICS-5000 HPLC system equipped with an AS-AP autosampler in a sequential injection configuration using the Chromeleon software version 7. 10 μl of the samples were injected on a 3 × 250 mm Dionex Carbopac PA200 column (Thermo Scientific, Waltham, United States). 56 μM of the β-1,3/1,4-MLG tetrasaccharide or 45 μM of the β-1,3/1,4-MLG trisaccharide were loaded onto the column. The gradient was used as follows: 0–5 min, 10% B, 3.5% C (initial conditions); 5–12 min 10% B, linear gradient from 0–30% C; 12.0–12.1 min, 50% B, 50% C; 12.1–13.0 min, exponential gradient of B and C, back to initial conditions, 13–17 min initial conditions. Solvent A was ultrapure water, solvent B was 1 M sodium hydroxide and solvent C was 1 M sodium acetate.
Matrix Assisted Laser Desorption Ionization–Time of Flight (MALDI-TOF) analysis of mixed-linkage glucans was performed with a Bruker Autoflex system (Bruker Daltonics) operated in reflectron mode. 10 mg/ml of the oligosaccharide were mixed 1:5 with 2,5-dihiydroxybenzoic acid in 1:1 H2O:MeOH on a Bruker MTP 384 grounded steel MALDI plate. The samples were allowed to dry and directly analyzed.
Barghahn S., Arnal G., Jain N., Petutschnig E., Brumer H, & Lipka V. (2021). Mixed Linkage β-1,3/1,4-Glucan Oligosaccharides Induce Defense Responses in Hordeum vulgare and Arabidopsis thaliana. Frontiers in Plant Science, 12, 682439.
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6

Oligosaccharide Characterization by HPAEC-PAD and MALDI-TOF

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High Performance Anion-Exchange Chromatography with Pulsed Amperometric Detection (HPAEC-PAD) was performed on a Dionex ICS-5000 DC HPLC system operated by the Chromeleon software version 7 (Dionex) using a Dionex Carbopac PA200 column. Solvent A was double-distilled water, solvent B was 1 M sodium hydroxide (NaOH), and solvent C was 1 M sodium acetate (NaOAc). The gradient used was: 0–4 min, 10% solvent B and 2.5% solvent C; 4–24 min, 10% B and a linear gradient from 2.5 to 25% C; 24–24.1 min, 50% B and 50% C; 24.1–25 min, an exponential gradient of NaOH and NaOAc back to initial conditions; and 25–31 min, initial conditions.
Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) was performed on a Bruker Daltonics Autoflex System (Billerica, USA). The matrix, 2,5-dihydroxy benzoic acid, was dissolved in 50% methanol in water to a final concentration of 10 mg mL−1. Oligosaccharide samples were mixed 1:1 (v/v) with the matrix solution. One microliter of this solution was placed on a Bruker MTP 384 ground steel MALDI plate and left to air dry for 2 h prior to analysis.
Attia M.A., Nelson C.E., Offen W.A., Jain N., Davies G.J., Gardner J.G, & Brumer H. (2018). In vitro and in vivo characterization of three Cellvibrio japonicus glycoside hydrolase family 5 members reveals potent xyloglucan backbone-cleaving functions. Biotechnology for Biofuels, 11, 45.
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7

N-Glycan Analysis of Rice Leaves

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N-glycans were prepared and purified from the mutant and wild-type rice leaves according to Karg et al. [55 (link)]. The N-glycan analysis was performed in positive-ion reflectron mode using a matrix-assisted laser desorption ionization-time of flight mass spectrometer (MALDI-TOF MS) with an Autoflex system from Bruker Daltonics (Bruker, Billerica, MA, USA). The tandem mass spectrometric analysis was carried out using an Axima Resonance MALDI-quadrupole ion trap-TOF instrument (Shimadzu, London, UK). 5-Dihydroxybenzoic acid (10 mg/mL in 50% methanol) was used as the matrix. The data analysis was performed with Flex Analysis 3.3 software (Bruker) and Launchpad 2.9.3 software (Kratos Analytical Ltd., Cambridge, UK). The calculated mass of the major N-glycans was compared with earlier reports to assign the corresponding structures [12 (link),13 (link),56 (link)].
Sim J.S., Kesawat M.S., Kumar M., Kim S.Y., Mani V., Subramanian P., Park S., Lee C.M., Kim S.R, & Hahn B.S. (2018). Lack of the α1,3-Fucosyltransferase Gene (Osfuct) Affects Anther Development and Pollen Viability in Rice. International Journal of Molecular Sciences, 19(4), 1225.
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