SEC-MALS analysis was performed with an HPLC system (Waters, Milford, MA, USA), coupled with UV- (Waters), Dawn8+ MALS- (Wyatt, Dernbach, Germany), and RI- (Shodex, Tokyo, Japan) detectors. Samples were filtered using 0.1 μm centrifugal filters (Millipore, Burlington, MA, USA) and injected onto Superdex 200 Increase 10/300 column (Cytiva, Marlborough, MA, USA), previously equilibrated with 20 mM Tris pH 7.5, 150 mM NaCl. Data were analyzed using Astra 7.0.
Dawn8 mals
Manufactured by Wyatt Technology
Sourced in Germany, United States
The DAWN8+ MALS is a multi-angle light scattering (MALS) detector designed for use in a variety of analytical applications. It measures the intensity of light scattered by macromolecules or particles in a sample at multiple angles, allowing for the determination of molecular weight, size, and shape.
Automatically generated - may contain errors
Lab products found in correlation
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Astra software, by Wyatt Technology (2 mentions)
Superdex 200 increase 10 300 column, by Cytiva (1 mentions)
Hplc system, by Waters Corporation (1 mentions)
Ultraviolet detector, by Shimadzu (1 mentions)
Uv detector, by Shimadzu (1 mentions)
Superose 12 10 300 gl column, by Cytiva (1 mentions)
3 protocols using dawn8 mals
1
SEC-MALS Analysis of Macromolecular Complexes
2
Characterizing Protein Complex Sizes
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The MWs of complexes were determined using 10/300 Superdex 200 pg increase columns (Cytiva) with inline DAWN8+ MALS (Wyatt Technology, Santa Barbara, CA, USA), an ultraviolet detector (Shimadzu, Tokyo, Japan), and a refractive index detector (Shodex, New York, NY, USA). Protein samples (45 μL) were injected at 20 μM bpAb with 20 μM MtsA. Analysis was performed using ASTRA software (Wyatt Technology). The protein concentration was calculated from the refractive index. All detectors were calibrated using bovine serum albumin (Sigma‐Aldrich, St. Louis, MO, USA).
3
Molecular Weight Determination of LI-Cadherin
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The MW of LI-cadherin was determined using Superose 12 10/300 GL column (Cytiva) with inline DAWN8+ MALS (Wyatt Technology), UV detector (Shimadzu), and refractive index detector (Shodex). Protein samples (45 μl) were injected at 100 μM or 50 μM. Analysis was performed using ASTRA software (Wyatt Technology). The concentration at the end of the chromatographic column was measured based on the UV absorbance. The protein conjugate method was used for the analysis as sugar chains were bound to LI-cadherin. All detectors were calibrated using bovine serum albumin (Sigma-Aldrich).
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