Purified samples were loaded onto a Superdex 75 10/300 GL column (GE) attached to an Äkta Prime chromatography system. The runs were performed at a flow rate of 0.5 mL/min using a degassed buffer made of 50 mM Tris–HCl pH 8.0, 100 mM NaCl. Molecular weight standards were used to calibrate the column (MWGF1000 kit for molecular weights 29,000–700,000 and apronitin, Sigma-Aldrich). The molecular weight of the protein ClpT1 was also determined on a Precision Detectors PD2010 light scattering instrument connected in tandem to a high-performance liquid chromatography system as described in [46 (link)].
Mwgf1000 kit
Manufactured by Merck Group
Sourced in United States
The MWGF1000 kit is a lab equipment product manufactured by the Merck Group. The kit is designed for DNA and RNA purification, providing a reliable and efficient solution for researchers and scientists. The core function of the MWGF1000 kit is to facilitate the extraction and purification of nucleic acids from various sample types. The product details and specifications can be obtained from the Merck Group's technical documentation.
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Lab products found in correlation
4 protocols using mwgf1000 kit
1
Molecular Weight Determination of ClpT1
2
Protein Purification via Size-Exclusion Chromatography
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Individual proteins (tRip‐N and ERS‐N) were injected onto a Superdex 200 Increase 10/300 column (GE Healthcare) while the chimeric protein and complexes (≥100 kDa) were purified on a SepFast SEC 6–5000 kDa column (BioToolomics). SEC columns were run with the same SEC buffer (25 mM HEPES‐NaOH pH 7.0, 300 mM NaCl, 5% (v/v) glycerol, 0.005% (w/v) DDM, 5 mM β‐ME) and were periodically calibrated either with Bio‐Rad's gel filtration standard (#1511901) or MWGF1000 kit (Sigma‐Aldrich), respectively, to determine molecular weight (MW) estimates from the chromatograms. In addition, blue dextran was used to determine the column's void volume (V0).
3
Molecular Mass Determination of IaaH
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The relative molecular mass of the native IaaH was determined by size exclusion chromatography. The Ni-NTA agarose beads purified enzyme (10 μg) was loaded onto a TSKgel G2000SWXL (125 Å, 5μm, 7.8× 300 mm) HPLC column (TOSOH Bioscience LLC, USA). The enzyme was eluted with 100 mM sodium phosphate buffer (pH 8.0) containing 100 mM sodium sulfate at a flow rate of 1 ml/min. The relative molecular mass was then calculated from the calibration curve obtained with standard proteins (MWGF1000 Kit, Sigma-Aldrich, Co., St. Louis, USA).
4
Protein Molecular Weight Determination
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HiLoad 16/60 Superdex 200 preparatory grade (GE Healthcare, Amersham) column was calibrated using a set of protein molecular weight markers (MWGF1000 kit; Sigma-Aldrich Chemicals, St. Louis, Missouri, USA). The column was equilibrated and then developed (1.0 ml min−1) at room temperature with the purification buffer containing 0.15 M NaCl. The elution volume (Ve) of blue dextran (2000 kDa) was taken as void volume (Vo). Elution coefficient (Kav) [28] was determined using the formula: Kav = (Ve-Vo)/(Vt-Vo), where Vt is the total column volume (120 ml).
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