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Size exclusive chromatography

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Size exclusive chromatography is a technique used for the separation and purification of molecules based on their size and molecular weight. It allows for the separation of compounds in a mixture by their ability to penetrate the porous matrix of the stationary phase within the chromatographic column.

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Lab products found in correlation

Protease inhibitor, by Merck Group (3 mentions) Microfluidizer, by Microfluidics (3 mentions)

3 protocols using size exclusive chromatography

1

Purification of Recombinant Proteins

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All recombinant proteins were expressed in E. coli BL21(DE3) codon plus and induced by 1.0 mM isopropyl thiogalactoside (IPTG) at 16°C overnight. The bacterial pellets were lysed using a microfluidizer (Microfluidics) in binding/ATPase buffer (100 mM Tris-HCl pH 7.5, 150 mM NaCl, 5 mM MgCl2, and 5% Glycerol) with protease inhibitors (Sigma) and centrifuged at 15,000 × g for 30 min. Histagged and GST-tagged proteins were purified by using Nickel-NTA and glutathione beads, respectively following manufacturer’s instructions. Protein samples eluted from the beads were further purified by size exclusive chromatography (GE Healthcare) using the ATPase buffer. The purity of the recombinant proteins was checked by Coomassie staining and western immunoblotting.
Umanah G.K., Ghasemi M., Yin X., Chang M., Kim J.W., Zhang J., Ma E., Scarffe L.A., Lee Y.I., Chen R., Tangella K., McNamara A., Abalde-Atristain L., Dar M.A., Bennett S., Cortes M., Andrabi S.A., Doulias P.T., Ischiropoulos H., Dawson T.M, & Dawson V.L. (2020). AMPA Receptor Surface Expression Is Regulated by S-Nitrosylation of Thorase and Transnitrosylation of NSF. Cell reports, 33(5), 108329.
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2

Purification of Recombinant Proteins from E. coli

Check if the same lab product or an alternative is used in the 5 most similar protocols
All recombinant proteins were expressed in E. coli BL21(DE3) codon plus and induced by 1.0 mM isopropyl thiogalactoside (IPTG) at 16 °C overnight. The bacterial pellets were lysed using a microfluidizer (Microfluidics) in binding/ATPase buffer (100 mM Tris-HCl pH 7.5, 150 mM NaCl, 5 mM MgCl2, and 5% Glycerol) with protease inhibitors (Sigma) and centrifuged at 15,000 × g for 30 min. GST-tagged proteins were purified by using glutathione beads, respectively following manufacturer’s instructions. Protein samples eluted from the beads were further purified by size exclusive chromatography (GE Healthcare) using the ATPase buffer. The purity of the recombinant proteins was checked by Coomassie staining and western immunoblotting.
Umanah G.K., Abalde-Atristain L., Khan M.R., Mitra J., Dar M.A., Chang M., Tangella K., McNamara A., Bennett S., Chen R., Aggarwal V., Cortes M., Worley P.F., Ha T., Dawson T.M, & Dawson V.L. (2022). AAA + ATPase Thorase inhibits mTOR signaling through the disassembly of the mTOR complex 1. Nature Communications, 13, 4836.
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3

Purification of Recombinant Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
All recombinant proteins were expressed in E. coli BL21(DE3) codon plus and induced by 1.0 mM isopropyl thiogalactoside (IPTG) at 16°C overnight. The bacterial pellets were lysed using a microfluidizer (Microfluidics) in binding/ATPase buffer (100 mM Tris-HCl pH 7.5, 150 mM NaCl, 5 mM MgCl2, and 5% Glycerol) with protease inhibitors (Sigma) and centrifuged at 15,000 × g for 30 min. Histagged and GST-tagged proteins were purified by using Nickel-NTA and glutathione beads, respectively following manufacturer’s instructions. Protein samples eluted from the beads were further purified by size exclusive chromatography (GE Healthcare) using the ATPase buffer. The purity of the recombinant proteins was checked by Coomassie staining and western immunoblotting.
Umanah G.K., Ghasemi M., Yin X., Chang M., Kim J.W., Zhang J., Ma E., Scarffe L.A., Lee Y.I., Chen R., Tangella K., McNamara A., Abalde-Atristain L., Dar M.A., Bennett S., Cortes M., Andrabi S.A., Doulias P.T., Ischiropoulos H., Dawson T.M, & Dawson V.L. (2020). AMPA Receptor Surface Expression Is Regulated by S-Nitrosylation of Thorase and Transnitrosylation of NSF. Cell reports, 33(5), 108329.
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