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Urine samples were tested for S. hematobium infection by using the Urine Filtration Kit (Vestergaard-Frandsen, Nairobi, Kenya) provided by WHO.
The gel filtration kit (GE-Healthcare) was used to calibrate a Superdex 200 column with high and low molecular weight standards, previously equilibrated with 20 mM MOPS buffer (pH 6.75) containing 0.1 M NaCl.
The column was calibrated using a High Molecular Weight Gel Filtration kit (GE Healthcare) containing the following standards: Blue Dextran 2000 (void volume determination), thyroglobulin (669 kDa, Rs = 85.0 Å), ferritin (440 kDa, Rs = 61.0 Å), aldolase (158 kDa, Rs = 48.1 Å), conalbumin (75 kDa, Rs = 36.4 Å), and ovalbumin (44 kDa, Rs = 30.5 Å).
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More than 200,000 people gained access to clean water as UNICEF and its partners delivered emergency water, installed pumps and distributed water-filtration kits.
Molecular mass standards used for calibration were from the Gel Filtration Calibration Kit HMW (GE Healthcare, UK).
Protein sizes were estimated based on a column calibration with a mixture of low molecular weight proteins (gel filtration calibration kit LMW; GE Healthcare).
The size of protein complexes in each fraction was calculated by using the High Molecular weight gel filtration calibration kit (GE Healthcare).
The molecular mass of MEP synthase and the mutant derivatives were estimated by loading 1 mg of protein onto a Sephacryl 200HR (Sigma Aldrich, St . Louis MO) size-exclusion chromatography column equilibrated with 0.1 M Tris pH 7.5, 1 mM NaCl, 5 mM DTT (flow rate of 2 mL/min) and calibrated with a gel filtration standard kit purchased from Bio-Rad (Hercules, CA).
A standard curve was obtained using a high-molecular mass gel filtration calibration kit (GE Healthcare).
A calibration curve was plotted using the standard proteins in a gel filtration calibration kit (GE Healthcare).
The column was calibrated with the low molecular weight gel filtration calibration kit (Amersham Biosciences).
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Justyna Jupowicz-Kozak
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