Sodium chloride, trizma, glycine, SDS, EDTA, ANS, Gdm-HCl, PMSF, and imidazole were obtained from Sigma Chemical Co. Nickel–nitrilotriacetate (Ni–NTA) agarose–based resin was brought from Qiagen. Mono-Q ion-exchange and superdex-75 (10/300 GL) gel filtration columns were purchased from GE Healthcare. LB agar, LB broth, glycerol, ammonium persulphate, IPTG, and kanamycin were purchased from Himedia Laboratories. Syringe filters (0.22 and 0.02 μm cutoff) were purchased from Millipore Corporation. All chemicals and reagents used were of analytical grade.
Mono q ion exchange
Manufactured by GE Healthcare
The Mono-Q ion-exchange column is a laboratory equipment used for the purification and separation of biomolecules, such as proteins, peptides, and nucleic acids. It functions by utilizing ion-exchange chromatography, a widely used technique in biochemistry and molecular biology research. The Mono-Q column is designed to effectively bind and elute target molecules based on their ionic interactions with the column's positively charged resin.
Automatically generated - may contain errors
Lab products found in correlation
Syringe filter, by Merck Group (1 mentions)
Ammonium persulphate, by HiMedia (1 mentions)
Imidazole, by Merck Group (1 mentions)
Trizma, by Merck Group (1 mentions)
Lb agar, by HiMedia (1 mentions)
Isopropyl β d 1 thiogalactopyranoside (iptg), by HiMedia (1 mentions)
Gdmhcl, by Merck Group (1 mentions)
Lb broth, by HiMedia (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Merck Group (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions)
Sodium chloride (nacl), by Merck Group (1 mentions)
Sodium dodecyl sulfate (sds), by Merck Group (1 mentions)
Glycine, by Merck Group (1 mentions)
Kanamycin, by HiMedia (1 mentions)
Glycerol, by HiMedia (1 mentions)
2 protocols using mono q ion exchange
1
Purification of Recombinant Proteins
2
Recombinant Human Cytosolic KRS Purification
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The cDNAs for human cytosolic KRS (1–597 aa WT and N-terminal 12 aa truncated) were subcloned into pET-28a (Novagen) with EcoRI and XhoI and expressed in Escherichia coli BL21 (DE3). His-tagged KRS was then purified using nickel affinity (Invitrogen) and Mono Q ion-exchange (GE Healthcare) chromatography following the manufacturer’s instructions. To remove lipopolysaccharide, the KRS solution was dialyzed in pyrogen-free buffer and subsequently filtered through Acrodisc Units with Mustang E membrane, (0.2 µm, 25 mm; Pall Corporation) in PBS containing 20% glycerol.
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