Kta upc 900 fplc system

Molecular weights of DNMT1, DNMT3A-3L, and DNMT3B-3L
were measured by SEC-MALS. Superdex Increase 200 10/300 columns (GE
Healthcare) connected to a DAWN HELIOS II-18 angle MALS (Wyatt Technology)
detector with wavelength set to 680 nm were equilibrated in 20 mM
Tris-HCl, at pH 8.0, 200 mM NaCl, and 1 mM TCEP with a flow rate of
0.2 mL/min using ÄKTA-UPC 900 FPLC system (GE Healthcare).
Purified protein samples were centrifuged at 17 000g for 15 min at 4 °C and filtered through a 0.22 μm
filter (Millipore). Protein samples (100 μL injection volumes)
were injected into Superdex Increase 200 10/300 columns. UV fluorescence,
MALS, and refractive index data were recorded and analyzed using ASTRA
software (Wyatt Technology).

Molecular weights of Suv3 and Suv3ΔC were measured by SEC‐MALS. S200 10/300 columns connected to a DAWN HELIOS II‐18 angle MALS (Wyatt Technology) detector with wavelength set at 658 nm were equilibrated in 25 mM HEPES pH 8.0, 300 mM KCl, and 2 mM DTT and with a flow rate of 0.2 mL/min using the ÄKTA‐UPC 900 FPLC system (GE Healthcare). Purified protein samples were centrifuged at high speed for 15 min at 4°C and filtered through a 0.22‐μM filter (Millipore). Protein samples (100 μL injection volumes) were injected into the Superdex S200 10/300 columns. UV fluorescence, MALS, and Refractive Index data were recorded and analyzed using ASTRA software (Wyatt Technology).

The molecular mass of wild‐type and mutated TDP‐43 RRM proteins was measured by SEC‐MALS (size exclusion chromatography coupled with multi‐angle light scattering). The purified protein samples were centrifuged at 4°C and 13,000 rpm for 30 min and filtered through a 0.22‐μm Millex‐GV filter (Millipore). The samples were injected into a pre‐equilibrated Agilent Bio SEC‐3100 Å column (Agilent Technologies, for RRM1) or Superdex 75 10/300 GL column (GE Healthcare, for RRM1‐RRM2) connected to a DAWN HELIOS II‐18 angle MALS instrument (Wyatt Technology) with a refractive index (RI) detector (Optilab T‐rEX, Wyatt Technology). Samples were run using the ÄKTA‐UPC 900 FPLC system (GE Healthcare). The UV, scattering and refractive index data were analyzed using ASTRA software (Wyatt Technology) to calculate the molar mass of each sample.