Size exclusion chromatography with multiangle light scattering (SEC-MALS) data were collected using an Agilent 1260 HPLC system (Agilent Technologies, Santa Clara, CA), attached to a WYATT DAWN HELEOS-II MALS instrument and a Wyatt Optilab rEX differential refractometer (Wyatt Technology, Santa Barbara, CA). For separation, the KRAS4b-FMe/CaM complex was loaded onto a superdex 200 10/300 column (GE Healthcare) in a buffer containing 20 mM Hepes (pH 7.2), 150 mM NaCl, 1 mM CaCl2, 1 mM MgCl2, and 1 mM TCEP. Data were analyzed using the ASTRA VI software (Wyatt Technology).
Wyatt optilab rex
Manufactured by Wyatt Technology
Sourced in United States
The Wyatt Optilab rEX is a multi-angle light scattering (MALS) detector that measures the absolute molar mass and size of macromolecules in solution. It provides precise and reliable data on the molecular weight and radius of gyration of proteins, polymers, and other large molecules.
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Lab products found in correlation
Pump 1100 series, by Agilent Technologies (2 mentions)
Dawn eos, by Wyatt Technology (2 mentions)
Superdex 200 10 300 column, by GE Healthcare (1 mentions)
Astra 6 software, by Wyatt Technology (1 mentions)
Agilent 1260, by Agilent Technologies (1 mentions)
Lc 20ad pump, by Shimadzu (1 mentions)
Sil 20a ht autosampler, by Shimadzu (1 mentions)
G1379a degasser, by Agilent Technologies (1 mentions)
Dawn 8, by Wyatt Technology (1 mentions)
Wtc 030s5 size exclusion column, by Wyatt Technology (1 mentions)
Akta purifier upc10 fplc protein purification system, by Wyatt Technology (1 mentions)
4 protocols using wyatt optilab rex
1
SEC-MALS Analysis of KRAS4b-FMe/CaM Complex
2
Molecular Weight Determination of Polymers
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The number average molecular weights (Mn) of PFGE and PFGE-b-PIGG were determined by using an Agilent Pump 1100 Series (preceded by an Agilent G1379A Degasser), equipped with a set of two PLGel 5 μMIXED-C columns. A Wyatt Optilab Rex differential refractometer and Dawn Eos (Wyatt Technology Corporation, Santa Barbara, CA, USA) laser photometer were used as detectors. Dichloromethane was used as eluent at a flow rate of 0.8 mL min−1 at room temperature.
The number average molecular weights (Mn) of PFGE-b-PGGE was determined by gel permeation chromatography (GPC) using a Shimadzu Pump LC-20AD and Shimadzu SIL-20A HT Autosampler. A refractometer RI-Optilab-T-rex-Wyatt and laser photometer DAWN 8+ (Wyatt Technology) were used as detectors. N,N′-dimethylformamide was used as eluent at a flow rate of 0.8 mL min−1 at 25 °C.
The number average molecular weights (Mn) of PFGE-b-PGGE was determined by gel permeation chromatography (GPC) using a Shimadzu Pump LC-20AD and Shimadzu SIL-20A HT Autosampler. A refractometer RI-Optilab-T-rex-Wyatt and laser photometer DAWN 8+ (Wyatt Technology) were used as detectors. N,N′-dimethylformamide was used as eluent at a flow rate of 0.8 mL min−1 at 25 °C.
3
Determination of KS Molecular Weight
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The molecular weight of KS was determined by static light scattering (SLS) using a Wyatt Dawn Heleos II multi-angle light scattering detector (Wyatt Technology). This was coupled to an AKTA Purifier UPC10 FPLC protein purification system equipped with a WTC-030S5 size-exclusion column (Wyatt Technologies). 2.5 mg/mL KS (100 μL) was applied to the size-exclusion column with a buffer containing 25 mM Tris (pH 7.5), 150 mM NaCl, 10 mM DTT and 0.02% NaN3 using a flow rate of 0.5 mL/min. BSA (2 mg/mL) was used for the system calibration. The molecular weights of the individual peaks in the size-exclusion chromatogram were determined from the SLS results in conjunction with refractive index measurements (Wyatt Optilab rEX, connected downstream of the LS detector). A standard value of the refractive index, dn/dc = 0.185 mL/g, was used for the proteins and the buffer viscosity η = 1.0408 cP at 25°C, was used. The value of the reference refractive index, 1.3462, was taken directly from the measurement of the Wyatt Optilab rEX when only buffer was passing through the reference cell.
4
Synthesis and Characterization of PLLA-g-CNT and LPSQ-COOMe
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Commercial PLLA Luminy® L175 was obtained from Total Corbion (The Netherlands) with D-lactide content below 1%, according to the supplier. Weight average molar mass Mw of 78 kg/mol and dispersity Mw/Mn of 1.8 were measured by size exclusion chromatography (SEC) with a multi-angle laser light scattering detector in dichloromethane using an Agilent Pump 1100 Series (preceded by an Agilent G1379A Degasser), equipped with a set of two PLGel 5 µ MIXED-C columns. A Wyatt Optilab Rex differential refractometer and a Dawn Eos (Wyatt Technology Corporation) laser photometer were used as detectors. Dichloromethane was used as eluent at a flow rate of 0.8 mL min-1 at room temperature (RT).
MWCNT Nanocyl NC-7000 (Nanocyl, Belgium) with an average diameter of 9.5 nm, length of 1.5 μm, and 90% purity, were used. PLLA covalently attached to MWCNT (PL-g-CNT) was synthesized by ring-opening polymerization of L,L-lactide according to a well-established procedure [53 (link),54 (link)]. LPSQ-COOMe was synthesized as previously described [38 (link),39 (link)]. Procedures for the synthesis of PL-g-CNT and LPSQ-COOMe are described below.
Dichloromethane purchased from StanLab (Poland) (purity 99.5%) and acetone purchased from POCH (Poland) (purity 99%) were used as received.
MWCNT Nanocyl NC-7000 (Nanocyl, Belgium) with an average diameter of 9.5 nm, length of 1.5 μm, and 90% purity, were used. PLLA covalently attached to MWCNT (PL-g-CNT) was synthesized by ring-opening polymerization of L,L-lactide according to a well-established procedure [53 (link),54 (link)]. LPSQ-COOMe was synthesized as previously described [38 (link),39 (link)]. Procedures for the synthesis of PL-g-CNT and LPSQ-COOMe are described below.
Dichloromethane purchased from StanLab (Poland) (purity 99.5%) and acetone purchased from POCH (Poland) (purity 99%) were used as received.
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