Phenyl sepharose 6 fast flow resin

Manufactured by GE Healthcare

Phenyl Sepharose 6 Fast Flow resin is a hydrophobic interaction chromatography (HIC) medium used for the purification of proteins and other biomolecules. It consists of highly cross-linked, rigid agarose beads with covalently coupled phenyl groups. This resin is designed for fast, efficient, and scalable purification processes.

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Lab products found in correlation

Econo column chromatography column, by Bio-Rad (2 mentions) Pre packed, by GE Healthcare (1 mentions) Sephadex g 25 pd 10 column, by GE Healthcare (1 mentions) Oligonucleotide primer, by Merck Group (1 mentions) Amicon stirred cell concentrators, by Merck Group (1 mentions) Ultrafiltration membranes 10 000 da mw cutoff, by Merck Group (1 mentions) 0.45 μm membrane, by Merck Group (1 mentions)

Close spelling variants of this product

Phenyl-Sepharose (11 citations) Phenyl Sepharose 6 Fast Flow (5 citations) Phenyl Sepharose Fast Flow (2 citations)

3 protocols using phenyl sepharose 6 fast flow resin

Synthesis and Purification of Muconate Analogs

Cited 2 times

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Chemicals, biochemicals, buffers, solvents, and the components for Luria-Bertani (LB) media were obtained from sources reported elsewhere [11 (link)]. The synthesis of 2-hydroxymuconate (3a) [8 (link)], 2-hydroxy-2,4-pentadienoate (5a) [10 (link)], 5-chloro-2-hydroxymuconate (4Z-3b) [11 (link)], and (4Z)-5-chloro-2-hydroxy-2,4-pentadienoate (5b) [11 (link)], and ethyl 2-fluorocrotonate [18 (link)] are reported in the indicated references. The Phenyl Sepharose 6 Fast Flow resin and the pre-packed PD-10 Sephadex G-25 columns were obtained from GE Healthcare (Piscataway, NJ). The Econo-Column chromatography columns were obtained from Bio-Rad Laboratories, Inc. (Hercules, CA). 4-OT, 4-OD/VPH, and 4-OD/E106QVPH (all from P. putida mt-2) were purified by procedures reported elsewhere with minor modifications [10 (link)–11 (link)19 (link)–20 (link)]. The YM-3 ultrafiltration membranes and centrifugal microconcentrators were obtained from Millipore (Billerica, MA). Activities were determined using previously described assays [10 (link)–11 (link)19 (link)–20 (link)]. The plasmids containing the genes for 4-OT and 4-OD/VPH from P. putida mt-2 and L. cholodnii SP-6, and 4-OD/E106QVPH from P. putida mt-2 were constructed as reported elsewhere [10 (link)–11 (link)]. Proteins were expressed as described.

Stack T.M., Jr. W.H, & Whitman C.P. (2017). Synthesis and enzymatic ketonization of the 5-(halo)-2-hydroxymuconates and 5-(halo)-2-hydroxy-2,4-pentadienoates. Beilstein Journal of Organic Chemistry, 13, 1022-1031.

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Purification and Analysis of Compound 10

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Chemicals, biochemicals, buffers, and solvents were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO), Fisher Scientific Inc. (Pittsburgh, PA), Fluka Chemical Corp. (Milwaukee, WI), or EMD Millipore, Inc. (Billerica, MA). 2-Carboxybenzaldehyde (10) was obtained from Sigma-Aldrich Chemical Co. Phenyl-Sepharose 6 Fast Flow resin was obtained from GE Healthcare Bio-sciences (Pittsburgh, PA). The Econo-Column^® chromatography columns were obtained from BioRad (Hercules, CA). The Amicon stirred cell concentrators and the ultrafiltration membranes (10,000 Da, MW cutoff) were purchased from EMD Millipore Inc. Oligonucleotide primers were synthesized by Sigma-Aldrich.

LeVieux J.A., Medellin B., Johnson WH J.r., Erwin K., Li W., Johnson I.A., Zhang Y.J, & Whitman C.P. (2018). Structural Characterization of the Hydratase-Aldolases, NahE and PhdJ: Implications for Specificity, Catalysis, and the N-acetylneuraminate lyase subgroup of the Aldolase Superfamily. Biochemistry, 57(25), 3524-3536.

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Secreted Protein Expression in Bacillus subtilis

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Example 1

DNA sequences encoding variant α-amylases, a signal peptide for secretion, and additional 5′ and 3′ sequences for amplification and subcloning were constructed by standard PCR amplification or ordered from a commercial vendor (GeneArt). Standard procedures were used to insert the DNA sequence into a bacterial vector for secreted protein expression in Bacillus subtilis cells. The constructs were verified by DNA sequencing. Cells were grown for about 68 hours in expression medium suitable for secreted protein expression from B. subtilis. Cells were separated from protein-containing supernatant by centrifugation followed by filtration through 0.45 μm membranes (EMD Millipore). When needed, additional purification was achieved through ion exchange chromatography with Phenyl Sepharose 6 Fast Flow resin (GE Healthcare). Protein concentration was determined by high performance liquid chromatography (HPLC) and absorbance at 280 nm.

US11441139B2. α-Amylase variants (2022-09-13). DANISCO US INC. [US]. Inventors: Jonathan K. Lassila [US], Sandra W. Ramer [US].

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