vertical slab gel electrophoresis apparatus  (Bio-Rad)

Journal: Marine Drugs

Article Title: Purified Pyropia yezoensis Pigment Extract-Based Tandem Dye Synthesis

doi: 10.3390/md22050197

Figure Lengend Snippet: Summary of red phycoerythrin (R-PE) extraction and purification methods reported in the literature.

Article Snippet: Polysiphonia urceolata , Thaw frozen algae in 0.02 M Na-phosphate buffer (pH 7.0), filter, and centrifuge to obtain the PE-containing supernatant. Fractionate supernatant with ammonium sulfate at 25% and 45% ( w / v ), collect red supernatant and precipitate, dissolve precipitate in 20 mM phosphate buffer (pH 7.0), and dialyze overnight. Apply dialyzed R-PE samples on DEAE SepharoseTM Fast Flow column pre-equilibrated with 20 mM phosphate buffer (pH 5.6) containing 0.05 M NaCl. Wash the column with the same buffer, elute with 20 mM phosphate buffer containing 0.05 M NaCl with a pH gradient (pH 5.6–4.0, 2 × 50 mL) at 1 mL·min −1 , monitor eluate at 280 nm, and collect 2-mL fractions. Perform native PAGE and SDS-PAGE using the Bio-Rad vertical slab discontinuous gel electrophoresis apparatus with 5% stacking gel as well as 7.5% and 15% separating gel, respectively. Stain gels with CBB R250. , 1.79 (ca.) , 5.6 , Liu et al. [ ] .

Techniques: Extraction, Purification, Algae, Ion Exchange Chromatography, Filtration, Chromatography, Fast Protein Liquid Chromatography, Spectrophotometry, Clear Native PAGE, Staining, Residue, Centrifugation, High Performance Liquid Chromatography, Spectroscopy, Lysis, Concentration Assay, Nucleic Acid Electrophoresis, Isolation, Fluorescence, Electrophoresis

Journal: Marine Drugs

Article Title: Purified Pyropia yezoensis Pigment Extract-Based Tandem Dye Synthesis

doi: 10.3390/md22050197

Figure Lengend Snippet: Analysis of crude and three-step-purified red phycoerythrin (R-PE) extracts from Pyropia yezoensis . ( a ) Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of crude and purified R-PE extracts. Lane M, protein marker; Lane PE crude , crude protein extract; Lane PE purified , three-step-purified R-PE; ( b ) High-performance liquid chromatography analysis to determine the purity of R-PE in the extracts before ( left ) and after ( right ) the three-step separation process.

Article Snippet: Polysiphonia urceolata , Thaw frozen algae in 0.02 M Na-phosphate buffer (pH 7.0), filter, and centrifuge to obtain the PE-containing supernatant. Fractionate supernatant with ammonium sulfate at 25% and 45% ( w / v ), collect red supernatant and precipitate, dissolve precipitate in 20 mM phosphate buffer (pH 7.0), and dialyze overnight. Apply dialyzed R-PE samples on DEAE SepharoseTM Fast Flow column pre-equilibrated with 20 mM phosphate buffer (pH 5.6) containing 0.05 M NaCl. Wash the column with the same buffer, elute with 20 mM phosphate buffer containing 0.05 M NaCl with a pH gradient (pH 5.6–4.0, 2 × 50 mL) at 1 mL·min −1 , monitor eluate at 280 nm, and collect 2-mL fractions. Perform native PAGE and SDS-PAGE using the Bio-Rad vertical slab discontinuous gel electrophoresis apparatus with 5% stacking gel as well as 7.5% and 15% separating gel, respectively. Stain gels with CBB R250. , 1.79 (ca.) , 5.6 , Liu et al. [ ] .

Techniques: Purification, Polyacrylamide Gel Electrophoresis, Marker, High Performance Liquid Chromatography