Omnisec 4

Manufactured by Malvern Panalytical

Sourced in United Kingdom

The OmniSEC 4.7 is a multi-detector size exclusion chromatography (SEC) system designed for the analysis of macromolecules and nanoparticles. It is capable of providing information about the molecular weight, size, and conformation of various samples.

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Lab products found in correlation

Cbm 20a, by Shimadzu (4 mentions) Labsolutions version 5, by Shimadzu (2 mentions) Omnisec software, by Malvern Panalytical (2 mentions) Labsolution version 5, by Shimadzu (2 mentions) Sil 20a, by Shimadzu (1 mentions) Superdex 200 increase 5 150 gl column, by GE Healthcare (1 mentions) Superdex 75 5 150 gl column, by GE Healthcare (1 mentions) Bovine serum albumin (bsa), by Malvern Panalytical (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Superdex 200 increase 5 150, by GE Healthcare (1 mentions) β amylase, by Merck Group (1 mentions) Superdex 200 5 150 column, by GE Healthcare (1 mentions) Viscotek 270 dual detector, by Malvern Panalytical (1 mentions)

6 protocols using omnisec 4

Oligomeric State Analysis of Antidin sbAvd-7

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The oligomeric state of the antidin sbAvd-7 was analyzed with size exclusion chromatography (SEC) using a liquid chromatography instrument (CBM-20A, Shimadzu Corporation) equipped with an autosampler (SIL-20A), UV-Vis (SDP-20A), and a fluorescence detector (RF-20Axs). The instrument was integrated with a static light scattering instrument (SLS, Zetasizer μV light scattering detector (Malvern Instruments Ltd.)) to determine molecular weight of the eluted proteins. The instrument was controlled using Lab Solutions Version 5.51 (Shimadzu Corporation) and OmniSEC 4.7 (Malvern Instruments Ltd.). Samples (~50 μg in 10–100 μl) were injected onto a Superdex200 Increase 5/150GL column (GE Healthcare) and equilibrated with the buffer the protein was dialyzed against (50 mM sodium phosphate, 650 mM NaCl, pH 7) with a flow rate of 0.1 ml/min at 20°C. Molecular weight determination was done by calculating a standard curve based on the elution volume of the molecular weight markers (CA, carbonic anhydrase 29 kDa; BSA, Bovine Serum Albumin 66 kDa; ADH, Alcohol Dehydrogenase 150 kDa; BA, ß-Amylase 200 kDa, Sigma-Aldrich), and alternatively, using the light-scattering intensity-based determination protocol involving BSA (monomeric peak) in SLS detector calibration using a Malvern microV detector and the OmniSEC software (Malvern Instruments Ltd.) (Fig 6).

Agrawal N., Lehtonen S.I., Uusi-Mäkelä M., Jain P., Viitala S., Määttä J.A., Kähkönen N., Azizi L., Riihimäki T.A., Kulomaa M.S., Johnson M.S., Hytönen V.P, & Airenne T.T. (2019). Molecular features of steroid-binding antidins and their use for assaying serum progesterone. PLoS ONE, 14(2), e0212339.

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Protein Characterization by Liquid Chromatography

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Proteins were analysed using a liquid chromatography instrument (CBM-20A, Shimadzu Corporation, Kyoto, Japan) equipped with autosampler (SIL-20A), UV-VIS (SPD-20A) and fluorescence detector (RF-20Axs) as well as Zetasizer µV light scattering detector (Malvern Instruments Ltd, Worcestershire, UK) for molecular weight (static light scattering) and hydrodynamic size (dynamic light scattering) determination. The instrument was controlled using Lab Solutions Version 5.51 (Shimadzu Corporation) and OmniSEC 4.7 (Malvern Instruments Ltd.). Samples (70 µg in 40–100 µl) were injected on a Superdex75 5/150GL column (GE healthcare, Uppsala, Sweden) equilibrated with Na₂HPO₄/NaH₂PO₄, 650 mM NaCl, pH 7. Runs were executed with a flow rate of 0.25 ml/min at 12°C. Molecular weight determination was either done by calculating a standard curve of molecular weight markers (cytochrome C, 12.4 kDa; carbonic anhydrase, 29 kDa; ovalbumin, 44 kDa; BSA, 66 kDa, Sigma-Aldrich) or was based on the light-scattering intensity of the eluting protein, for which BSA was used for instrument calibration using the OmniSEC software (Malvern Instruments Ltd.).

Taskinen B., Airenne T.T., Jänis J., Rahikainen R., Johnson M.S., Kulomaa M.S, & Hytönen V.P. (2014). A Novel Chimeric Avidin with Increased Thermal Stability Using DNA Shuffling. PLoS ONE, 9(3), e92058.

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Protein Characterization by Liquid Chromatography

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Proteins were analyzed using a liquid chromatography instrument (CBM-20A; Shimadzu Corporation, Kyoto, Japan) equipped with an autosampler (SIL-20A), ultraviolet-visible (UV-VIS) (SPD-20A), and a fluorescence detector (RF-20Axs). Molecular weight was determined using SEC with a Malvern Zetasizer µV instrument (Malvern Instruments Ltd.) measuring inline SLS and DLS. Data were processed using Lab Solution Version 5.51 (Shimadzu Corporation) and OmniSec 4.7 (Malvern Instruments Ltd.) software. Samples (50 µg) were injected into the column (Superdex 200 Increase 5/150; GE Healthcare, Uppsala, Sweden) using the autosampler. The column was equilibrated with 50 mM Na₃PO₄, 150 mM NaCl, pH 7.2 running buffer. The measurements were run with a flow rate of 0.1 mL/min at 20 °C. The monomeric peak of bovine serum albumin (BSA) was used for calibrating the system to calculate molecular weight from the measured SLS intensity.

Zhang P., Azizi L., Kukkurainen S., Gao T., Baikoghli M., Jacquier M.C., Sun Y., Määttä J.A., Cheng R.H., Wehrle-Haller B., Hytönen V.P, & Wu J. (2020). Crystal structure of the FERM-folded talin head reveals the determinants for integrin binding. Proceedings of the National Academy of Sciences of the United States of America, 117(51), 32402-32412.

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Molecular Weight Determination of Zebrafish CA VI-PTX

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Molecular weight determination of zebrafish CA VI–PTX was performed using a Malvern Zetasizer μV instrument (Malvern Instruments Ltd., Worcestershire, UK) running static light scattering (SLS) and dynamic light scattering (DLS) methods. Analysis was performed using a liquid chromatography instrument (CBM-20A, Shimadzu Corporation, Kyoto, Japan) equipped with autosampler (SIL-20A), UV–VIS (SPD-20A) and fluorescence detector (RF-20Axs). Data were processed using Lab Solution Version 5.51 (Shimadzu Corporation) and OmniSec 4.7 (Malvern Instruments Ltd., Worcestershire, UK) softwares. A sample of the protein (50 μg) was injected on a Superdex 200 5/150 column (GE Healthcare, Uppsala, Sweden) equilibrated with 50 mM NaH₃PO₄, 500 mM NaCl pH 8 buffer. Runs were performed with flow rate of 0.1 ml/min at 20 °C using a thermostated cabin. The MW of the zebrafish CA VI–PTX was determined either by using a standard curve based on MW standard proteins (SEC analysis; CA 29 kDa, alcohol dehydrogenase 150 kDa, β-amylase 200 kDa, BSA 66 kDa, Sigma-Aldrich, Inc., St. Louis, MO, USA) or by calibrating the light scattering detector using the monomeric peak of BSA and light-scattering intensity (SLS).

Patrikainen M.S., Tolvanen M.E., Aspatwar A., Barker H.R., Ortutay C., Jänis J., Laitaoja M., Hytönen V.P., Azizi L., Manandhar P., Jáger E., Vullo D., Kukkurainen S., Hilvo M., Supuran C.T, & Parkkila S. (2017). Identification and characterization of a novel zebrafish (Danio rerio) pentraxin–carbonic anhydrase. PeerJ, 5, e4128.

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Molecular Weight Analysis of WSF

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Three milligrams of lyophilized WSF was suspended in 1 mL of water and added to 5 µL of LiOH 0.1 M. The mixture was filtered using a Whatman 0.45 µm PVDF filter (Grosseron, Coueron, France) and submitted to HPSEC (high-pressure size exclusion chromatography) analysis with refractive index (Viscotek VE 3580 RI detector, Malvern Instruments, Malvern, UK), multi-angle laser light scattering (low angle 7° and right angle) and viscosimetric detections (both via Viscotek 270 dual detector, Malvern Instruments). The samples were injected through a Shodex OHpak 805KB column equipped with an OHPak SB-G guard column (Thermo Fisher Scientific, Illirch, France). Pullulans were used as standards for system calibration. Molecular weight was obtained using the Omnisec 4.7 software (Malvern Instruments).

Palmont D., Bonnin E., Smith Ravin E.J., Lahaye M, & Marcelin O. (2024). Characterization of Crude and Processed Pulp Cell Walls of Three Selected Mamey Accessions (Mammea americana L.). Molecules, 29(7), 1596.

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Molecular Weight Determination of PNVCL-COOH

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Gel permeation chromatography (GPC) was performed on a Viscotek 270 model dual detector (Malvern Instruments, Malvern, UK), with A3000 Aq and A4000 Aq type columns. Tetrahydrofuran (THF) was the preferred solvent, and was introduced with a flow rate of 1 mL/min at 35.0 C. The number average (M n ) and weight average (Mw) molecular weights of PNVCL-COOH were measured by evaluating the GPC data using OmniSEC 4.7 software (Malvern Instruments, Malvern, UK).

Durkut S, & Elçin Y.M. (2017). Synthesis and characterization of thermosensitive poly(N-vinylcaprolactam)-g-collagen. Artificial cells, nanomedicine, and biotechnology, 45(8).

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