Superose 6 10 300 gl column

Manufactured by GE Healthcare

Sourced in United States, Sweden, United Kingdom, Germany, Ireland

The Superose 6 10/300 GL column is a size exclusion chromatography column designed for the purification and analysis of macromolecules. It is suitable for the separation and fractionation of proteins, peptides, nucleic acids, and other biomolecules based on their molecular size and shape.

Automatically generated - may contain errors

Lab products found in correlation

Vitrobot mark 4, by Thermo Fisher Scientific (13 mentions) Akta purifier, by GE Healthcare (8 mentions) Superdex 200 10 300 gl column, by GE Healthcare (7 mentions) Infinity reagent, by Thermo Fisher Scientific (6 mentions) Biologic duoflow quadtec 10 system, by Bio-Rad (5 mentions) Superdex 200, by GE Healthcare (4 mentions) Gel filtration calibration kit, by GE Healthcare (4 mentions) Pelco easiglow, by Ted Pella (4 mentions) Gel filtration standard, by Bio-Rad (4 mentions) Vivaflow tangential filtration cassettes, by Sartorius (3 mentions) Streptactin superflow column, by GE Healthcare (3 mentions) Effectene, by Qiagen (3 mentions) Hplc system, by Shimadzu (3 mentions) Akta purifier system, by GE Healthcare (3 mentions) Blue dextran 2000, by GE Healthcare (3 mentions) Amplex red cholesterol assay kit, by Thermo Fisher Scientific (3 mentions) Mwgf1000, by Merck Group (3 mentions) Astra 6, by Wyatt Technology (3 mentions) Ultimate 3000 hplc system, by Thermo Fisher Scientific (2 mentions) Atpγs, by Merck Group (2 mentions)

Spelling variants of this product

Superose 6 column (200 citations) Superose 6 Increase 10/300 GL column (191 citations) Superose 6 (82 citations) Superose 6 10/300 column (79 citations) Superose 6 10/300 GL (62 citations) Superose 6 10/300 (36 citations) Superose 6 10/300 GL SEC column (5 citations) Superose 6 10/300 GL prepacked column (3 citations) Superose 6 increase 10/300 GL SEC column (3 citations) 10/300 GL Superose 6 column (2 citations)

247 protocols using superose 6 10 300 gl column

Plasma Lipoprotein Fractionation and Analysis

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

Plasma samples were pooled (n = 2–4) and diluted 1:3.2 with PBS containing 0.01% EDTA and 500 μL were run at a Superose 6 10/300 GL column (17-5172-01, GE Healthcare). Fractions of 6 drops (∼280 μL) were collected. Protein profiles were determined by the absorbance read at 280 nm and cholesterol determined in individual fractions. Pools corresponding to the VLDL, LDL, and HDL peaks were collected for WB analysis of apoM.
Conditioned cell medium from respectively the apical (diluted 1:3) and the basolateral compartment was concentrated x6 using spin columns (10 k, Umicon Ultra, Merck) and pooled (medium from 3 wells). 500 μL of each sample were run at a Superose 6 10/300 GL column (17-5172-01, GE Healthcare) and fractions of 3 drops (∼140 μL) were collected. Also, a human plasma sample was run as reference material, and cholesterol determined in the fractions.

Bisgaard L.S., Christensen P.M., Oh J., Torta F., Füchtbauer E.M., Nielsen L.B, & Christoffersen C. (2024). Kidney derived apolipoprotein M and its role in acute kidney injury. Frontiers in Pharmacology, 15, 1328259.

+ Open protocol

+ Expand

Solubilization and Fractionation of ATP13A2

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

The membrane fractions of ATP13A2-transfected HEK293 cells and SH-SY5Y cells were solubilized in lysis buffer supplemented with 1.5% DDM and 0.3% CHS. Insoluble material was removed by ultracentrifugation at 366,000 × g, 30 min at 4 °C. The samples were then injected into a Superose 6 10/300 GL column (GE Life Sciences) connected to the AKTAexplorer 10 XT FPLC system (GE Healthcare) at 4 °C, equilibrated in a running buffer containing 50 mM Tris pH 7.5, 150 mM NaCl, 0.03% DDM, and 0.0015% CHS. Fractions of 1 mL were collected, precipitated with 10% trichloroacetic acid (TCA), and analyzed by Western blotting.

Fujii T., Nagamori S., Wiriyasermkul P., Zheng S., Yago A., Shimizu T., Tabuchi Y., Okumura T., Fujii T., Takeshima H, & Sakai H. (2023). Parkinson’s disease-associated ATP13A2/PARK9 functions as a lysosomal H+,K+-ATPase. Nature Communications, 14, 2174.

+ Open protocol

+ Expand

Fractionation of Cyanobacterial Proteome

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

Cells from vegetative cells or heterocysts were suspended in lysis buffer containing 20 mM Tris-Cl (pH = 7.5), 150 mM NaCl, 1% dodecylmaltoside, and Complete Protease Inhibitors EDTA-free (Roche), and sonicated on ice with an output of 135 W. The whole-cell lysate was centrifuged (10,000g at 4 °C for 10 min) to remove the cell debris. The protein concentration was determined using the Bradford assay. Then, 300 μg of protein mixtures of vegetative cells and heterocysts were individually fractionated by size-exclusion chromatography (SEC) using a Thermo Scientific Ultimate 3000 HPLC system. The lysates separated by SEC were injected into a Superose 6 10/300GL column (GE Life Sciences) equilibrated with PBS (pH = 7.2) and exposed to 120 min of isocratic elution. The total collection time was 55 min, as the first fraction was collected at 20 min and the last fraction finished at 75 min. In total, 55,300 μl fractions were collected, with a flow rate of 0.3 ml min⁻¹.

Xu C., Wang B., Heng H., Huang J, & Wan C. (2022). Comparative Network Biology Discovers Protein Complexes That Underline Cellular Differentiation in Anabaena sp.. Molecular & Cellular Proteomics : MCP, 21(4), 100224.

+ Open protocol

+ Expand

Membrane Protein Purification Protocol

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

Endogenously tagged MBRL-mNG HeLa cells were expanded to 3x 15 cm dishes. Cells were harvested and resuspended in 60 mL of chilled PBS (Sigma-Aldrich) supplemented with a cOmplete protease inhibitor cocktail table (Roche), 1 mM EDTA, 1 mM PMSF (Roche), 1.5 μM pepstatin A (Sigma-Aldrich) and 20 μg/mL of DNase I (Roche). Lysis was performed by passing the cells 5 times through an Avestin EmulsiFlex-C5 at 12,000 PSI of backpressure. The lysate was centrifuged at 12,000 g for 20 min at 4°C and the supernatant collected. Membranes were pelleted by centrifugation at 100,000 g for 60 min at 4°C. Pelleted membranes were resuspended in 2 mL of resuspension buffer containing 100 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1% DDM (Anatrace), 0.1% CHS (Anatrace), 1 mM EDTA, 1 mM PMSF and 1.5 μM pepstatin A. Membranes were solubilised for 2 h on a rotator at 4°C and insoluble material pelleted by centrifugation at 100,000 g for 45 min at 4°C. Solubilised material was applied to a 24 mL Superose 6 10/300 GL column (GE Healthcare Life Sciences) equilibrated with 2 column volumes of 50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 0.03% DDM and 0.003% CHS. Elutions were fractioned by 1.0 mL and aliquots run by SDS-PAGE for western blotting.

van de Weijer M.L., Krshnan L., Liberatori S., Guerrero E.N., Robson-Tull J., Hahn L., Lebbink R.J., Wiertz E.J., Fischer R., Ebner D, & Carvalho P. (2020). Quality Control of ER Membrane Proteins by the RNF185/Membralin Ubiquitin Ligase Complex. Molecular Cell, 79(5), 768-781.e7.

+ Open protocol

+ Expand

Characterization of Francisella Lipopolysaccharide Oligosaccharides

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

SEC profiles of hydrolyzed OAgs extracted under different hydrolytic conditions from F. tularensis LPS were run on a Sephacryl S-300 HR column (GE Healthcare Life Sciences) at 0.5 mL⋅min⁻¹ in PBS (pH 7.4). The average molecular weight was calculated with a dextran calibration curve (Sigma–Aldrich).
LMW, HMW, and VHMW OAgs extracted from F. tularensis LVS were run on a Superose 6 10/300 GL column (GE Healthcare Life Sciences) at 0.5 mL⋅min⁻¹ in PBS (pH 7.4). The average molecular weight was calculated with a dextran calibration curve (Sigma–Aldrich).
SEC analysis was also used to characterize conjugates, comparing them with free OAg and free TT. All samples were eluted on two Superose 6 10/300 GL columns connected in series for a better separation of conjugate from free saccharide and protein. The mobile phase consisted of PBS (pH 7.4) at 0.5 mL⋅min⁻¹. Void- and bed-volume calibrations were performed with λ-DNA (λ-DNA Molecular Weight Marker III, 0.12–21.2 Kbp; Roche) and sodium azide (NaN₃; Merck), respectively. OAg peaks were detected by a dRI, while UV detection at 214 nm and 280 nm was used for free protein and conjugate. For K_d determination, the following equation was used: K_d = (Te − T0)/(Tt − T0), where Te is the elution time of the analyte, T0 is the elution time of the biggest fragment of λ-DNA, and Tt is the elution time of NaN₃.

Stefanetti G., Okan N., Fink A., Gardner E, & Kasper D.L. (2019). Glycoconjugate vaccine using a genetically modified O antigen induces protective antibodies to Francisella tularensis. Proceedings of the National Academy of Sciences of the United States of America, 116(14), 7062-7070.

+ Open protocol

+ Expand

Purification of PARP Proteins from E. coli

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

HIS-tagged human PARP1, PARP2, GST-tagged human PARP10, and GST-tagged human PARP1 catalytic domain (GST-PARP1 CAT, aa. 662–1014) proteins were expressed in Escherichia coli BL21. Cells were grown in LB media and induced with 200 μM isopropyl 1-thio-β-d-galactopyranoside at 16 °C for 20 h. Proteins fused to GST were purified using glutathione-Sepharose beads according to the manufacturer’s protocols (GE Healthcare). HIS-tagged proteins were purified by Ni Sepharose 6 Fast Flow according to the manufacturer’s instruction (GE Healthcare). All recombinant proteins were further purified by passing through Superose 6 10/300 GL column (GE Healthcare) in 50 mM sodium phosphate buffer, pH 7.0, and 150 mM NaCl. Expression and purification of all recombinant proteins was analyzed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) followed by Coomassie staining.

Bian C., Zhang C., Luo T., Vyas A., Chen S.H., Liu C., Kassab M.A., Yang Y., Kong M, & Yu X. (2019). NADP+ is an endogenous PARP inhibitor in DNA damage response and tumor suppression. Nature Communications, 10, 693.

+ Open protocol

+ Expand

Plasma Lipoprotein Profiling by FPLC

Cited 1 time

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

Plasma lipoprotein profile was analyzed by FPLC as described ^{42 (link)}. Briefly, after 100 μl plasma was injected, lipoproteins were run at 0.5 ml/min in a buffer containing 0.15 M NaCl, 0.01 M Na₂HPO4, 0.1 mM EDTA, pH 7.5, and separated on a Superose 6 10/300 GL column (GE Healthcare) by using BioLogic DuoFlow QuadTec 10 System (Bio-Rad, CA). 500 μl of sample per fraction was collected. Triglyceride or cholesterol levels in each fraction were determined using Infinity reagents (Thermo Fisher).

Li Y., Xu Y., Jadhav K., Zhu Y., Yin L, & Zhang Y. (2019). Hepatic forkhead box protein A3 regulates ApoA-I expression, cholesterol efflux and atherogenesis. Arteriosclerosis, thrombosis, and vascular biology, 39(8), 1574-1587.

+ Open protocol

+ Expand

High-resolution FPLC analysis of HDL and LPS conjugates

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

HDL and 594-HDL-488-LPS conjugates were analyzed using high resolution FPLC on Superose 6 Increase 3.2/300 column and preparatively-purified on Superose 6 10/300 GL column (both from GE Healthcare Life Sciences). The FPLC setup consisted of Bio-Rad DuoFlow equipped with C96 autosampler with cooling, and BioFrac fraction collector (all from Bio-Rad). Lipoproteins were eluted at a flow rate of 0.05 mL/min in all analytical FPLC runs and 0.5 mL/min in all preparative runs. All lipoproteins were eluted isocratically using 10 mM phosphate buffer and 154 mM NaCl at pH 7.4. Fractions were collected by automatic fraction collector at 4°C (Bio-Frac) in 1.2 ml sized micro-titer tubes, followed by analysis of fluorescence and absorbance on the plate reader (Molecular Devices SpectraMax i3). FPLC chromatograms were then plotted as fluorescence intensity (or absorbance) against fraction number.

Yao Z., Mates J.M., Cheplowitz A.M., Hammer L.P., Maiseyeu A., Phillips G.S., Wewers M.D., Rajaram M.V., Robinson J.M., Anderson C.L, & Ganesan L.P. (2016). Blood-borne LPS is rapidly eliminated by liver sinusoidal endothelial cells via HDL. Journal of immunology (Baltimore, Md. : 1950), 197(6), 2390-2399.

+ Open protocol

+ Expand

Molecular Weight Determination of NarGH Complexes

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

A Superose 6 10/300 GL column (GE Healthcare) was equilibrated with 10 CV of Tris-HCl 40 mM pH 7.6, 8% glycerol buffer using an ÄKTA purifier fast-protein liquid chromatography (FPLC) machine (GE Healthcare). Molecular weight calibration of the column was done by using the following standard markers as recommended by the manufacturer (Sigma-Aldrich): blue dextran (2000 kDa), thyroglobulin (660 kDa), apoferritin (443 kDa), β-amylase (200 kDa) and alcohol dehydrogenase (150 kda). Holo, apo and R108A NarGH complexes were injected at a flow rate of 0.3 ml/min, protein elution being monitored at 280 nm. Varying protein concentration did not influence the elution profile.

Arias-Cartin R., Ceccaldi P., Schoepp-Cothenet B., Frick K., Blanc J.M., Guigliarelli B., Walburger A., Grimaldi S., Friedrich T., Receveur-Brechot V, & Magalon A. (2016). Redox cofactors insertion in prokaryotic molybdoenzymes occurs via a conserved folding mechanism. Scientific Reports, 6, 37743.

+ Open protocol

+ Expand

Structural Analysis of CtLas1-Grc3 Complex

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

Purified CtLas1-Grc3 complex (10 μM) was cross-linked with 50 μM bis(sulfosuccinimidyl)suberate (BS3; Sigma) in 20 mM Hepes pH 7.7, 200 mM NaCl, 5 mM MgCl₂, 5% glycerol at room temperature for 5 minutes before quenching with 30 mM Tris pH 7.5 for 15 minutes at 4°C. Cross-linked reactions were resolved over a Superose 6 10/300 GL column (GE Healthcare) in 10 mM Tris pH 8.0, 200 mM NaCl, 5 mM MgCl₂. CtLas1-Grc3 (0.15 mg/ml) was incubated in the absence (apo state) and presence (ATP-γS bound state) of 2 mM ATP-γS (Sigma) and 10 μM CT-ITS2-RNA (5′-UGUGUUGGGGdeoxyACCCGCGGCUGCUCG CGGGCCCUGAAAAGCA-3′) for 1 hour at 4°C. The CT-ITS2-RNA substrate represents part of the C. thermophilum ITS2 pre-rRNA sequence. Protein mixtures (3 μL) were applied to glow-discharged UltrAuFoil R1.2/1.3 300 mesh grids (Quantifoil) and vitrified after 5 sec blotting using the Automatic Plunge Freezer EM GP (Leica).

Pillon M.C., Hsu A.L., Krahn J.M., Williams J.G., Goslen K.H., Sobhany M., Borgnia M.J, & Stanley R.E. (2019). Cryo-EM Reveals Active Site Coordination Within a Multienzyme pre-rRNA Processing Complex. Nature structural & molecular biology, 26(9), 830-839.

+ 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!