Aeris widepore c4 column

Manufactured by Phenomenex

The Aeris Widepore C4 column is a reversed-phase high-performance liquid chromatography (HPLC) column designed for the separation and analysis of large biomolecules. It features a wide pore structure that facilitates the efficient separation of proteins, peptides, and other macromolecules.

Automatically generated - may contain errors

Lab products found in correlation

Synapt g2 si, by Waters Corporation (4 mentions) Dionex ultimate 3000 hplc, by Thermo Fisher Scientific (2 mentions) Zspray esi source, by Waters Corporation (2 mentions) 1200 series system, by Agilent Technologies (1 mentions) Chemstation software, by Agilent Technologies (1 mentions) Luna c18 2 column, by Phenomenex (1 mentions) Bio toolkit, by AB Sciex (1 mentions) Dionex ultimate 3000 hplc system, by Thermo Fisher Scientific (1 mentions) Tripletof 5600, by AB Sciex (1 mentions) Masslynx v4, by Waters Corporation (1 mentions) Chromeleon 6, by Thermo Fisher Scientific (1 mentions) Ultimate 3000 lc system, by Thermo Fisher Scientific (1 mentions) Dataanalysis version 4, by Bruker (1 mentions) Maxis 2 electrospray mass spectrometer, by Bruker (1 mentions) Synapt g2 si q tof mass spectrometer, by Waters Corporation (1 mentions) Picotip emitter, by New Objective (1 mentions) Ultimate 3000 hplc system, by Thermo Fisher Scientific (1 mentions)

8 protocols using aeris widepore c4 column

Analytical HPLC of Pharmaceutical Compounds

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

Analytical HPLC was carried out on an Agilent 1200 series system with UV detection at 214 nm. Methods used are summarized as follows, referenced by number in the text and in figures:
HPLC Method 1: Column - Phenomenex Aeris Widepore C4 column, 150 × 4.6 mm, 3.6 μm, 200 Åsilica; flow rate 0.8 mL/minute; Solvent System - A = water with 0.1% TFA, B = acetonitrile with 0.08% TFA; Gradient − 3 minute hold 1% B, 1–61% B gradient over 60 minutes, 3 minute hold 61% B, 10-minute post run 1% B; Flow Rate − 0.8 mL/minute
HPLC Method 1a: As in Method 1, but with a 1–61% B gradient over 30 minutes.
HPLC Method 2: Column - Phenomenex Luna C18(2 ) column, 100 × 4.6 mm, 3 μm, 100 Å silica; flow rate 1.0 mL/minute; Solvent System - A = water with 0.1% TFA, B = acetonitrile with 0.08% TFA; Gradient - 3 minute hold 1% B, 1–61% B gradient over 60 minutes, 3 minute hold 61% B, 10-minute post run 1% B
Integrals of HPLC peaks were calculated automatically with Agilent ChemStation software with subsequent manual inspection of the magnified baseline and modification of the automated calls - most commonly removal of erroneous peaks more consistent with background variation or splitting of a major peak to reflect tailing.

Albin J.S, & Pentelute B.L. (2020). Efficient flow synthesis of human antimicrobial peptides. Australian journal of chemistry, 73(4), 380-388.

+ Open protocol

+ Expand

Acyl-FosACP Enzymatic Assay Protocol

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

The above-described samples of acyl-FosACP were divided into 2 equal 100–150 μL portions. To one, FosDH1 or FosDH2 was added to a final concentration of 50 μM while the blank was supplemented with an equivalent volume of 50 mM sodium phosphate, pH 7.5. After parallel 60-min incubations at room temperature, each reaction mixture was diluted with formic acid/H₂O and centrifuged for 5 min at 14000g. These samples were then analyzed by LC-ESI(+)-MS and LC-ESI(+)-MS/MS using an analytical Aeris widepore-C4 column (3.6 μm, 2.1 × 150 mm) from Phenomenex using a linear gradient from 30% to 70% CH₃CN/H₂O on a Thermo-LXQ mass spectrometer. For LC-ESI(+)-MS/MS analysis^{15 (link)} the M¹¹⁺ ion was selected for MS/MSs such that, both the hydrated and dehydrated pPant ejection fragments could be observed together (Figures S12, S13, S23, S24, S20, S21, and S27–S30).

Shah D.D., You Y.O, & Cane D.E. (2017). Stereospecific Formation of E- and Z- Disubstituted Double Bonds by Dehydratase Domains from Modules 1 and 2 of the Fostriecin Polyketide Synthase. Journal of the American Chemical Society, 139(40), 14322-14330.

+ Open protocol

+ Expand

Protein Separation and Mass Spectrometry

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

500 ng protein was dissolved in 20 mM Tris 7.6, 50 mM KCl, 1 mM TCEP and loaded on an Eksigent MicroLC column (ChromXP C4, 3 μm particle size, 300 Å pore diameter, dimensions 50 × 0.5 mm) using a Dionex Ultimate 3000 HPLC system (Thermo Scientific). The proteins were separated on a gradient from 5% to 90% acetonitrile in water and 0.1% formic acid for 30 min at a flow rate of 30 µL/min. Mass spectra were recorded on a Sciex TripleTOF 5600 instrument equipped with a Turbo V DuoSpray ion source in positive mode. Data was analysed in PeakView version 2.1 (Sciex) using the Bio Tool Kit to reconstruct the uncharged average protein mass. All other samples were diluted in 0.1% formic acid (FA) to a concentration of 10 ng/μL and 50 ng were loaded on an Aeris Widepore C4 column, 3.6 µm particle size, dimensions 2.1 × 150 mm (Phenomenex), using a Dionex Ultimate 3000 HPLC system (Thermo Scientific) with a working temperature of 55 °C, 0.1% FA as solvent A, 90% acetonitrile, 0.08% FA as solvent B. Proteins were separated in a 6 min gradient from 10 to 70% solvent B at a flow rate of 300 μL/min. Mass spectra were recorded on a Waters Synapt G2-Si equipped with a ZSpray ESI source. Data were analysed in MassLynx V 4.1 using the MaxEnt 1 process to reconstruct the uncharged average protein mass.

von Raußendorf F., de Ruiter A, & Leonard T.A. (2017). A switch in nucleotide affinity governs activation of the Src and Tec family kinases. Scientific Reports, 7, 17405.

+ Open protocol

+ Expand

LC-MS Analysis of Recombinant aIF5A Proteins

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

10 µl (0.5 mg/ml) of aIF5A-C-His, produced in Sso PH1-16, was analyzed by LC-MS in order to assess its intact mass and the eventual presence of post-translational modifications. The recombinant protein N-His-aIF5A produced in E. coli was analyzed and served as a control. High performance liquid chromatography was performed on a Dionex Ultimate 3000 HPLC (Thermo Fisher Scientific) system configured with the Chromeleon 6.0 software (Thermo Fisher Scientific). The proteins were reduced in 100 mM DTT for 30 min at room temperature and then separated on an Aeris Widepore C4 column (3.6 µm particle size, dimensions 2.1 × 150 mm, Phenomenex) running a 6 min step gradient from 10% up to 70% acetonitrile in 0.1% formic acid. The working temperature was set to 50 °C and the flow rate at 300 µl/min. The LC-system was coupled online to the quadrupole-time of flight-mass spectrometer Synapt G2-Si (Waters), operated via the MassLynx V 4.1 software package, using a Z Spray ESI source (Waters). Mass spectra were acquired in the m/z range from 500 to 2000 at a scan rate of 1 s and the mass spectrometer was calibrated with a MS spectrum of [Glu1]-Fibrinopeptide B human (Glu-Fib) solution. The data were analyzed with the MaxEnt algorithm to reconstruct the uncharged average protein mass.

Bassani F., Romagnoli A., Cacciamani T., Amici A., Benelli D., Londei P., Märtens B., Bläsi U, & La Teana A. (2018). Modification of translation factor aIF5A from Sulfolobus solfataricus. Extremophiles, 22(5), 769-780.

+ Open protocol

+ Expand

Intact Protein Mass Spectrometry Protocol

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

For intact protein MS, proteins were diluted in 0.1% formic acid (FA) to a concentration of 20 ng/μl, and 100 ng were loaded on an Aeris Widepore C4 column, 3.6-μm particle size, dimensions 2.1 × 150 mm (Phenomenex), using a Dionex Ultimate 3000 HPLC system (Thermo Scientific) with a working temperature of 55 °C: 0.1% FA as solvent A; 90% acetonitrile, 0.08% FA as solvent B. The proteins were separated on a 6-min gradient from 10 to 70% solvent B at a flow rate of 300 μl/min. Mass spectra were recorded on a Waters Synapt G2-Si equipped with a ZSpray ESI source. Glu[1]-Fibrinopeptide B (Glu-Fib) was used as a lock mass, and spectra were corrected on the fly. Data were analyzed in MassLynx version 4.1 using the MaxEnt 1 process to reconstruct the uncharged average protein mass.
The experimental details of peptide MS and phosphopeptide mapping of PKD1 and PKD1 quantification by parallel reaction monitoring are described in the supporting Experimental procedures.

Elsner D.J., Siess K.M., Gossenreiter T., Hartl M, & Leonard T.A. (2019). A ubiquitin-like domain controls protein kinase D dimerization and activation by trans-autophosphorylation. The Journal of Biological Chemistry, 294(39), 14422-14441.

+ Open protocol

+ Expand

Disulfide Bond Analysis of Chlorophyllase

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

To obtain information regarding the state of the disulfide bonds in solution, purified chlorophyllase and its variants were explored using intact mass spectrometry experiments. To accomplish this task, chlorophyllase and its variants were diluted to a final concentration of 10 μM in 20 mM Tris–HCl (pH 8.0) and assessed using LC-MS. Samples that were alkylated were incubated at 30 °C for 45 min with 10 mM iodoacetamide, except for C220S and C264S which were incubated with 2 mM iodoacetamide. For samples containing DTT, tris(2-carboxyethyl)phosphine, or GSH, 10 μM of WT chlorophyllase was incubated with 20 mM reductant for 30 min at 30 °C before injection onto the instrument. All LC-MS experiments were conducted using the instrumentation described above. An Aeris WIDEPORE C4 column (2.1 × 50 mm, 3.6 μm, 200 Å) (Phenomenex) was used for sample separations, with solvent A = water with 0.1% formic acid and solvent B = 95% acetonitrile, 5% water, and 0.1% formic acid. Each injection was 2 μl of sample and method used was run at 0.3 ml/min. The method consisted of three steps: i) 5% solvent B from 0 to 2 min, ii) a linear gradient flow to 80% solvent B for 5 min, iii) and finally an isocratic flow of 80% solvent B for 2 min. Results were analyzed in the Agilent MassHunter Bioconfirm software (https://www.agilent.com/).

Jo M., Knapp M., Boggs D.G., Brimberry M., Donnan P.H, & Bridwell-Rabb J. (2023). A structure-function analysis of chlorophyllase reveals a mechanism for activity regulation dependent on disulfide bonds. The Journal of Biological Chemistry, 299(3), 102958.

+ Open protocol

+ Expand

Intact Molecular Weight Analysis of Purified Enzyme

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

Example 2

Intact molecular weight analyses of the purified enzyme was performed using a MAXIS II elec-trospray mass spectrometer (Bruker Daltonik GmbH, Bremen, DE). The samples were diluted to 0.1 mg/ml in MQ water. The diluted sample was applied to an Aeris Widepore C4 column (Phenom-enex). The sample was washed and eluted from the column running an acetonitrile linear gradient and introduced to the electrospray source with a flow of 300 ml/min by an Ultimate 3000 LC system (Dionex). Data analysis is performed with DataAnalysis version 4.2 (Bruker Daltonik GmbH, Bremen, DE). The molecular weight of the sample was calculated by Maximum Entropy deconvolution of the raw data in the range 10.000 to 40.000 Da. The dominant peak observed at molecular weight of 28873.08 Da corresponds to within 0.3 Da of the calculated molecular weight of the mature protease in SEQ ID: 2.

US11001821B2. Polypeptides having protease activity and polynucleotides encoding same (2021-05-11). NOVOZYMES A/S [DK]. Inventors: Astrid Benie [DK], Morten Gjermansen [DK], Peter Rahbek Oestergaard [DK].

+ Open protocol

+ Expand

Protein Mass Determination by Q-ToF MS

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

The protein mass was determined on a Synapt G2-Si Q-ToF mass spectrometer (Waters) coupled to an Ultimate 3000 HPLC system (Dionex, Thermo Fisher Scientific) via a ZSpray ESI source (Waters). Proteins were separated on an Aeris Widepore C4 column (3.6 µm particle size, dimensions 2.1 x 150 mm, Phenomenex) with a working temperature of 55 °C, applying a 10 min gradient from 8 % to 63 % acetonitrile in 0.1 % formic acid at a flow rate of 300 µL/min. On the mass spectrometer the sampling cone voltage was 40 V, the desolvation gas temperature 450 ˚C and the source temperature 120 ˚C.
For MS experiments under native conditions, the buffer of the protein solutions was exchanged to 50 mM ammonium acetate, pH 6.8. Measurements were performed on the Synapt G2-Si equipped with the NanoLockSpray electrospray source (Waters) using pre-opened PicoTip emitters (New Objective). Electrospray voltage was set at 1.8 kV, the source temperature at 80 ˚C and sampling cone voltage at 80 V. All data were analysed in MassLynx V 4.1 using the MaxEnt 1 process to reconstruct the uncharged average protein mass.

Truebestein L., Waltenberger E., Gehin C., Gavin A., & Leonard T.A. (2022). Structure and regulation of the myotonic dystrophy kinase-related Cdc42-binding kinase.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!