Sepharose cl 6b

Manufactured by Merck Group

Sourced in United States

Sepharose CL-6B is a crosslinked agarose gel bead matrix used for size-exclusion chromatography. It is designed for the separation and purification of various biomolecules, such as proteins, nucleic acids, and polysaccharides, based on their molecular size.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

Sepharose 6B (22 citations) Sepharose CL-6B column (5 citations) DEAE-Sepharose CL-6B (5 citations) Sepharose CL-6B beads (3 citations) DEAE-Sepharose CL-6B column (2 citations)

11 protocols using sepharose cl 6b

Extraction and Characterization of Medicinal Plant Pectins

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

DEAE-cellulose, Sepharose CL-6B, Sephadex G25, and aminobenzamide (2AB) were purchased from Sigma-Aldrich. Endo-polygalacturonase (Endo-PG, EC 3.2.1.15 from Aspergillus niger) was purchased from Megazyme. All chemicals used were analytical grade and produced in China.
Homogalacturonan pectins were extracted from the following plants: Panax japonicus, Pseudostellaria heterophylla, Schisandra chinensis, Prunella vulgaris, Panax Notoginseng, Polygonum orientale, Anemarrhena asphodeloides, Kadsura longipedunculata, Isatis indigotica, Aconitum carmichaelii, Coptis chinensis, and Sophora flavescens.

Yu Y., Cui L., Liu X., Wang Y., Song C., Pak U., Mayo K.H., Sun L, & Zhou Y. (2022). Determining Methyl-Esterification Patterns in Plant-Derived Homogalacturonan Pectins. Frontiers in Nutrition, 9, 925050.

+ Open protocol

+ Expand

Characterization of Tobacco Leaf Polysaccharides

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

The flue-cured tobacco leaves (Yunyan 87) were kindly donated from the RandD center of Shanghai Tobacco (Group) Co., Ltd. (Shanghai, China). The tobacco leaves were dried at 45°C and ground into powder prior to the experiments. The standard monosaccharides including rhamnose, fucose, arabinase, xylose, mannose, galactose, glucose, glucuronic acid and galacturonic acid, 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and Sepharose CL-6B were obtained from Sigma (St. Louis, MO, USA). All other reagents/chemicals used were of analytical grade.

Xu C., Yang C, & Mao D. (2014). Fraction and chemical analysis of antioxidant active polysaccharide isolated from flue-cured tobacco leaves. Pharmacognosy Magazine, 10(37), 66-69.

+ Open protocol

+ Expand

Reagents for Cell Viability Assay

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

Reduced GSH, 1-chloro-2,4-dinitrobenzene (CDNB), ampicillin, sodium dodecyl sulfate (SDS), and the chromatographic material Sepharose CL-6B were purchased from Sigma-Aldrich, St. Louis, MO, USA (Merck, Rahway, NJ, USA) and were used without further treatment. Ethanol, methanol, and dimethyl sulfoxide (DMSO) were purchased from Scharlau (Barcelona, Spain). DU-145 culture media were obtained from Thermo Fisher Scientific (Waltham, MA, USA). The (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) reagent (MTT) was purchased from Applichem (Darmstadt, Germany).

Pantiora P., Furlan V., Matiadis D., Mavroidi B., Perperopoulou F., Papageorgiou A.C., Sagnou M., Bren U., Pelecanou M, & Labrou N.E. (2022). Monocarbonyl Curcumin Analogues as Potent Inhibitors against Human Glutathione Transferase P1-1. Antioxidants, 12(1), 63.

+ Open protocol

+ Expand

Synthesis and Characterization of CFL Lipid Probes

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

N-[N-(3-Triethoxysilyl)propylsuccinamoly]dihexadecylamine (i.e., CFL) was synthesized according to a previously reported method [18 ]. An N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamide (NBD-DPPE), an Alexa Flour® 488 goat anti-mouse IgG, Measure-IT™ thiol assay kit, and ProLong® gold antifade reagent with DAPI and LIVE/DEAD® viability/cytotoxicity kit for mammalian cells were purchased from Invitrogen (Carlsbad, CA), and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[maleimide(polyethylene glycol)-2000] (DSPE-PEG (2000) Maleimide) was from Avanti Polar Lipids (Alabaster, AL). Anti-EGFR (Ab-3) mouse mAb (225) was from EMD Millipore (Billerica, MA). Cholesterol and thiazolyl blue tetrazolium bromide (MTT) were from Alfa Aesar (Ward Hill, MA). Traut’s reagent (2-Iminothiolane•HCl) was from Thermo Scientific (Waltham, MA). A Bio-Rad protein assay kit was from Bio-Rad (Hercules, CA). Zeba spin desalting columns and Sepharose® CL-6B were from Sigma-Aldrich (St. Louis, MO). The A43 and HL-60 cell lines were purchased from ATCC (Manassas, VA). The DU-145 cell line was a generous gift from Dr. Daruka Mahadevan (Arizona Cancer Center, Univ. of Arizona).

Leung S.L., Zha Z., Cohn C., Dai Z, & Wu X. (2014). Anti-EGFR Antibody Conjugated Organic-Inorganic Hybrid Lipid Nanovesicles Selectively Target Tumor Cells. Colloids and surfaces. B, Biointerfaces, 121, 141-149.

+ Open protocol

+ Expand

Isolation and Characterization of Fibrinolytic Enzyme

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

Roots (100 mg/mL)
were kept overnight in sodium phosphate buffer (pH 7.5) at 4 °C.
Roots were cut, macerated, and centrifuged. The supernatant served
as the extract.
Isolation of the fibrinolytic enzyme was achieved
with substrate affinity chromatography (fibrinogen-coupled Sepharose
CL-6B, Sigma-Aldrich) followed by sodium dodecyl sulfate poly-acrylamide
gel electrophoresis (SDS-PAGE). Further purification involved size-exclusion
HPLC (SE-HPLC, Waters) with a Waters Protein-Pak 300 column pre-equilibrated
with 10 mM sodium phosphate buffer containing 0.1 M NaCl operated
at 0.8 mL/min. HPLC was performed on a Waters 600 HPLC system with
a Waters 2487 dual λ absorbance detector. A calibration curve
was constructed using marker proteins, and a linear dependence of
log M_w (molecular weight) versus
retention time was observed (R² = 0.9755,
where R² is the regression coefficient).
The approximate molecular weight of the enzyme was calculated from
the corresponding retention time that revealed the highest (major)
peak corresponding to 13.6 kDa (Figure S1, indicated by the area within dashed lines). Purification was further
confirmed with fibrinogen zymography.

Bhattacharyya R., Bhattacharjee S., Pathak B.K, & Sengupta J. (2020). Heptameric Peptide Interferes with Amyloid-β Aggregation by Structural Reorganization of the Toxic Oligomers. ACS Omega, 5(26), 16128-16138.

+ Open protocol

+ Expand

Protein Quantification and Purification

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

Sucrose, Tris-HCl, EDTA, NaOH, HCl, NaCl, isopropanol, glycine, cyanuric acid, 2,4,6-trinitrobenzenesulphonic acid (TNBS), 2-methylbutylamine, 4-aminobenzoic acid, sodium citrate, bicinchoninic acid (BCA), bovine serum albumin (BSA), p-nitrophenylbutyrate (p-NPB), acetonitrile, Sodium dodecyl sulfate, acrylamide and bia-acrylamide, ethanol, acetic acid, sodium thiosulfate, silver nitrate, formaldehyde, and sodium carbonate were purchased from Sigma-Aldrich (Darmstadt, Germany). BCA^TM (bicinchoninic acid) and Micro BCA^TM Protein Assays Reagents were from ThermoScientific (Rockford, IL, USA). Sepharose CL-6B was purchased from Sigma-Aldrich. The anion-exchange chromatography (DEAE-cellulose and Q-Sepharose) columns used were from GE Healthcare (Amersham, UK).

Fonseca L.P, & Taipa M.Â. (2024). One-Step Purification of Recombinant Cutinase from an E. coli Extract Using a Stabilizing Triazine-Scaffolded Synthetic Affinity Ligand. Biomimetics, 9(1), 57.

+ Open protocol

+ Expand

Enzymatic Synthesis and Purification of Fructan

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

Fructan, a component of NTM048 EPS, was synthesized by recombinant enzyme LvnS expressed in E. coli as described in our earlier study.^{27 (link)} The reaction was conducted in 50 mM sucrose, 1 mM CaCl₂, and 100 mM sodium acetate (pH 5.0) using the recombinant LvnS at 28°C for 24 h. The fructan produced was purified using Sepharose CL6B (Sigma–Aldrich Corp.), and freeze-dried. The absence of proteins in the fructan sample was confirmed by measuring the absorbance at 280 nm.

Matsuzaki C., Nakashima Y., Endo I., Tomabechi Y., Higashimura Y., Itonori S., Hosomi K., Kunisawa J., Yamamoto K, & Hisa K. (2021). Enzymatically synthesized exopolysaccharide of a probiotic strain Leuconostoc mesenteroides NTM048 shows adjuvant activity to promote IgA antibody responses. Gut Microbes, 13(1), 1949097.

+ Open protocol

+ Expand

Biochemical Analysis of Protein Complexes

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

Anti-Erg6, Ubx2, and Ldb16 were homemade antibodies that have been described previously (Wang and Lee, 2012 (link); Wang et al., 2014 (link)). The anti-S tag antibody (ab24838, mouse monoclonal) was from Abcam, anti-Act1 (actin) antibody (MAB1501, mouse monoclonal) was from Millipore, peroxidase anti-peroxidase (323-005-024, rabbit polyclonal) was from Jackson ImmunoResearch Laboratories, and anti-Flag antibody (F3165, mouse monoclonal) was from Sigma-Aldrich. Myriocin was from Sigma-Aldrich, and AbA was from Takara. IgG Sepharose 6 Fast Flow was from GE, Sepharose CL-6B was from Sigma-Aldrich, AcTEV protease was from Invitrogen, the Pierce silver staining kit was from Thermo Fisher Scientific, NADPH was from Sigma-Aldrich, palmitoyl-CoA was from Avanti Polar Lipids, and [2-¹⁴C] malonyl-CoA and [³H]-serine were from PerkinElmer. Complete EDTA-free protease inhibitor cocktail was from Roche. Phos-tag was from Wako Pure Chemical Industries. Bodipy 493/503 was from Invitrogen. All other common chemical reagents were from Sigma-Aldrich. Restriction and modifying enzymes for cloning and PCR were from Thermo Fisher Scientific.

Su W.C., Lin Y.H., Pagac M, & Wang C.W. (2019). Seipin negatively regulates sphingolipid production at the ER–LD contact site. The Journal of Cell Biology, 218(11), 3663-3680.

+ Open protocol

+ Expand

Ultrashort Carbon Nanotubes Synthesis and Characterization

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

All of the lipids (DOPC, 1-oleoyl-2-6-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]hexanoyl-sn-glycero-3-phosphocholine [NBD-PC], and cholesterol) were obtained from Avanti. All of the other chemicals were purchased from Sigma-Aldrich and used as received, unless specified. Live/dead assay and MTT cellular assay kits were obtained from Abcam. The size-exclusion columns for LUV separation used Sepharose CL-6B (Sigma-Aldrich). The ultrashort CNTPs were synthesized by sonication-assisted cutting of 0.8-nm single-walled carbon nanotubes according to the previously published procedure (33 (link)). Previous studies have confirmed that this procedure produces CNTPs with an extremely tight diameter distribution of 0.81

\pm

0.14 nm as measured by transmission electron microscopy (23 (link)). Some CNTP batches were chemically coupled to the 6-aminofluorescein (6-AF) dye using a 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) coupling procedure based on a published protocol (34 (link)). All fluorescence and absorbance spectra were measured with the Spectramax iD3 Microplate Reader (Molecular Devices) and Cytation 5 (Biotek). Vesicle size was measured using a dynamic light scattering (DLS) setup (Malvern Analytical).

Ho N.T., Siggel M., Camacho K.V., Bhaskara R.M., Hicks J.M., Yao Y.C., Zhang Y., Köfinger J., Hummer G, & Noy A. (2021). Membrane fusion and drug delivery with carbon nanotube porins. Proceedings of the National Academy of Sciences of the United States of America, 118(19), e2016974118.

+ Open protocol

+ Expand

Extraction and Purification of NTM048 EPS

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

As described in our earlier study,^{16 (link)} NTM048 EPS was extracted and purified. Briefly, 500 µl of an overnight bacterial culture of strain NTM048 was used to inoculate 50 ml of EPS production medium containing 15% sucrose, 0.5% bacto-peptone, 0.5% yeast extract, 1.5% K₂HPO₄, 0.001% MnCl₂·H₂O, 0.001% NaCl, and 0.005% CaCl₂, and the culture fluid was incubated for 24 h at 30°C. After the microorganisms were removed by centrifugation, the culture supernatant was precipitated by the addition of an equal volume of chilled ethanol, shaken vigorously, and centrifuged at 8,000 × g for 10 min. The supernatant was decanted, and this step was repeated twice. The precipitated EPS was then moderately sonicated to dissolve it in distilled water, followed to purification using Sepharose CL6B (Sigma–Aldrich Corp.) and freeze-drying. The absence of proteins in the EPS sample was confirmed by measuring the absorbance at 280 nm.

+ 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!