Sepharose beads

Manufactured by Qiagen

Sourced in United States

Sepharose beads are a type of cross-linked agarose-based chromatography medium used for the purification and separation of biomolecules, such as proteins, enzymes, and nucleic acids. They provide a high surface area for ligand immobilization and efficient adsorption and desorption of target molecules.

Automatically generated - may contain errors

Lab products found in correlation

Gelcode blue stain reagent, by Thermo Fisher Scientific (1 mentions) Supersignal west dura extended duration substrate, by Thermo Fisher Scientific (1 mentions) Anti pgk1 antibody, by Thermo Fisher Scientific (1 mentions) Cl xposure, by Thermo Fisher Scientific (1 mentions) Quikchange 2 xl site directed mutagenesis kit, by Agilent Technologies (1 mentions) Pgem t vector, by Promega (1 mentions) Pet28 vector, by Merck Group (1 mentions) Isopropyl β d 1 thiogalactopyranoside iptg, by Merck Group (1 mentions) Nickel nta, by Qiagen (1 mentions)

Spelling variants of this product

Glutathione sepharose beads (3 citations) Streptavidin-coated sepharose beads (3 citations) GSH-Sepharose beads (2 citations) Streptavidin Sepharose High Performance beads (2 citations) Streptavidin-sepharose beads (2 citations)

4 protocols using sepharose beads

Production and Purification of Recombinant Carp Cytokines

Check if the same lab product or an alternative is used in the 5 most similar protocols

Recombinant carp interferon gamma 2 (Ifn-γ) was produced as described previously^{15 (link)}. Protein analysis by SDS-PAGE (12% Tris–HCl, Bio-Rad) stained with GelCode Blue Stain Reagent (Thermo Scientific) revealed proteins were at least 95% pure and the chromogenic Limulus amebocyte lysate end-point test (Charles River Laboratories) showed that the residual endotoxin content was below detection limit (< 0.15 EU).
Recombinant carp Il-4/13b1 (previously named Il-4/13B) was produced essentially as described previously^{90 (link)} and the expression plasmid^{90 (link)} a kind gift of Professor T. Moritomo and Dr. F. Katakura, Laboratory of Comparative Immunology, Nihon University. In short, the poly-His-tagged Il-4/13b1 protein was expressed in Rosetta-gami B (DE3) pLysS Competent cells (Novagen) and purified using sepharose beads (Qiagen) followed by gel chromatography size exclusion using Superdex 200 Prep Grade 26/600 column (GE Healthcare). Protein analysis by SDS-PAGE (12% Tris–HCl, Bio-Rad) stained with GelCode Blue Stain Reagent (Thermo Scientific) revealed that proteins were at least 95% pure and residual endotoxin content was shown to be < 0.005 EU/ml (EndoZyme II Recombinant Factor C (rFC) Assay, Hyglos GmbH).

Wentzel A.S., Petit J., van Veen W.G., Fink I.R., Scheer M.H., Piazzon M.C., Forlenza M., Spaink H.P, & Wiegertjes G.F. (2020). Transcriptome sequencing supports a conservation of macrophage polarization in fish. Scientific Reports, 10, 13470.

+ Open protocol

+ Expand

Affinity Purification of Ubiquitin Conjugates

Check if the same lab product or an alternative is used in the 5 most similar protocols

Ubiquitylated proteins were purified from 50 OD₆₀₀ equivalents of exponentially growing yeast cells expressing 6His-tagged ubiquitin as previously described³³. 6His-ubiquitin conjugates were retained on nickel–nitrilotriacetic acid Sepharose beads (Qiagen) and eluted (eluate, E) in the presence of 300 mM (Stp1-HA, Stp1-RI_17-33-HA) or 500 mM (Stp2-HA, Stp2Δ_2-13-HA) imidazole. Total extracts (T), flow through (F) and eluate fractions were precipitated with 10% trichloroacetic acid, analysed by SDS-PAGE and immunoblotting with antibodies against the HA tag (1:5,000, Roche) and the signals were recorded using autoradiographic film (CL-Xposure, Thermo Scientific). As controls, levels of ubiquitin conjugates and Pgk1 were assessed with anti-His₅ (1:5,000, Qiagen) and anti-Pgk1 antibodies (1:10,000, In Vitrogen), respectively, and detected by chemiluminescence using SuperSignal West Dura Extended Duration Substrate (Thermo Scientific) and a Molecular Imager ChemDoc XRS+ with Image Lab v3 build 11 software (BioRad). Loaded T and F fractions correspond to 2% (Stp1-HA or Stp1-RI_17-33-HA) and 0.7% (Stp2-HA or Stp2Δ_2-13-HA) of the amount used for purification of ubiquitin conjugates.

Khmelinskii A., Blaszczak E., Pantazopoulou M., Fischer B., Omnus D.J., Le Dez G., Brossard A., Gunnarsson A., Barry J.D., Meurer M., Kirrmaier D., Boone C., Huber W., Rabut G., Ljungdahl P.O, & Knop M. (2014). Protein quality control at the inner nuclear membrane. Nature, 516(7531), 410-413.

+ Open protocol

+ Expand

Recombinant SelT Protein Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

Rat SelT cDNA devoid of signal peptide sequence was amplified from PC12 cells and cloned in the pGEM-T vector (Promega). Sequence mutagenesis was conducted by using the QuikChange^® II XL site-directed mutagenesis kit (Agilent Technologies) to delete the putative transmembrane domain and to convert the Cys-X-X-Sec motive to Cys-X-X-Cys or Ser-X-X-Ser (Fig. 1). The nucleotide sequence of all constructs was verified by DNA sequencing (Beckman Coulter Genomics). The different cDNAs were then subcloned in the pET28 vector (Millipore) for protein expression in BL21 bacteria. Protein synthesis was induced by adding 1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG; Sigma-Aldrich). Bacterial homogenates were centrifuged and incubated for 1 h with equilibrated sepharose beads coupled to Ni-nitroacetic acid (Qiagen). The beads were washed and proteins were eluted in the presence of 250 mM imidazole following the manufacturer's recommendations. Enzyme activity of recombinant SelT was assessed using thioredoxin reductase and glutathione peroxidase assay kits (Interchim).

Boukhzar L., Hamieh A., Cartier D., Tanguy Y., Alsharif I., Castex M., Arabo A., Hajji S.E., Bonnet J.J., Errami M., Falluel-Morel A., Chagraoui A., Lihrmann I, & Anouar Y. (2016). Selenoprotein T Exerts an Essential Oxidoreductase Activity That Protects Dopaminergic Neurons in Mouse Models of Parkinson's Disease. Antioxidants & Redox Signaling, 24(11), 557-574.

+ Open protocol

+ Expand

Purification and analysis of hydantoin compounds

Check if the same lab product or an alternative is used in the 5 most similar protocols

Chemicals were obtained from Sigma Aldrich and Thermo Fisher Scientific (USA). Nickel NTA and sepharose beads were purchased from Qiagen (USA). All enzymes were provided by New England BioLabs (USA). Lipids were purchased from Avanti Polar Lipids (USA) and detergents from Anatrace (USA). The hydantoin compounds were a kind gift of Marta Sans, Maria Kokkinidou and Arwen Pearson (University of Hamburg).

Majd H., King M.S., Palmer S.M., Smith A.C., Elbourne L.D., Paulsen I.T., Sharples D., Henderson P.J, & Kunji E.R. (2018). Screening of candidate substrates and coupling ions of transporters by thermostability shift assays. eLife, 7, e38821.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!