For each sample, 274 g of algal fronds were homogenized with 25 mM Tris-HCl (pH 7.2) and centrifuged at 15,000 g for 15 min. Ammonium sulfate was added to the supernatant to yield a 100% saturated solution and proteins were precipitated by centrifugation. The pellets were resuspended in 25 mM Tris-HCl buffer (pH 7.2) and dialyzed overnight against 25 mM Tris-HCl buffer (pH 7.2) to remove salt. The dialyzed samples were subjected to anion exchange chromatography on DEAE Sepharose FF (250 ml, Amersham Biosciences) and the column was eluted with 0.17 M, 0.27 M, and 0.37 M NaCl in Tris-HCl buffer (pH 7.2). The 0.27 M NaCl fraction, which had antibacterial activity, was subjected to gel filtration chromatography using Toyopearl HW-65S resin (particle size 20–40 μm, Tosoh), and the column was eluted with 25 mM Tris-HCl buffer (pH 7.2) at a 24 ml/min flow rate.
Toyopearl hw 65s resin
Manufactured by Tosoh
Toyopearl HW-65S resin is a size exclusion chromatography medium designed for the separation and purification of biomolecules such as proteins, peptides, and nucleic acids. It features a spherical porous structure with a controlled pore size distribution.
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Lab products found in correlation
2 protocols using toyopearl hw 65s resin
1
Purification of Antibacterial Proteins from Algae
2
Solid-Phase Oligonucleotide Synthesis Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Bead functionalization and solid-phase phosphoramidite oligonucleotide synthesis was performed by ATDBio. Toyopearl HW-65s resin, purchased from Tosoh Biosciences (product number: 0019815), was used as the solid support. Prior to oligonucleotide synthesis, the initial loading of hydroxyl groups on the resin was reduced via a capping reaction. Capping was performed by suspending the resin in a mixture of acetic anhydride and lutidine in THF, and N-methyl imadazole in acetonitrile for 24 hours. Following capping, the synthesis was performed using an ABI394 DNA synthesiser. The sequence of the capture oligonucleotide is
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