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Monos 10 100 column

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The MonoS 10/100 column is a laboratory equipment designed for protein purification. It is a strong cation exchange chromatography column that can be used to separate and purify proteins based on their charge. The column has a bed volume of 10 mL and can handle sample volumes up to 100 mL.

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Lab products found in correlation

Sf9 insect cells, by Expression Systems (3 mentions) Superdex 200 10 300 gl column, by GE Healthcare (3 mentions)

3 protocols using monos 10 100 column

1

Purification of Recombinant Human GRK5

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Full length human GRK5 was modified with a C-terminal hexa-histidine tag and cloned into pVL1392 vector for baculovirus production. GRK5 was expressed and purified as previously published (Beyett et al., 2019 (link)). Briefly, Sf9 insect cells (Expression Systems) were infected with a BestBac-derived baculovirus at a density of 3.5 × 106 cells/mL and harvested 48 hours post infection. Cells were resuspended, lysed by sonication and the supernatant was applied to Ni-NTA resin. The resin was washed with lysis buffer and GRK5 eluted with lysis buffer supplemented with 200 mM imidazole. The combined eluate was then subjected to cation-exchange chromatography using a MonoS 10/100 column (GE healthcare) and eluted with a linear gradient of NaCl. Fractions containing GRK5 were combined and run on a Superdex 200 10/300 GL column (GE healthcare). GRK5 was aliquoted, flash frozen, and stored at −80 °C until use.
Janetzko J., Kise R., Barsi-Rhyne B., Siepe D.H., Heydenreich F.M., Kawakami K., Masureel M., Maeda S., Garcia K.C., von Zastrow M., Inoue A, & Kobilka B.K. (2022). Membrane phosphoinositides regulate GPCR-β-arrestin complex assembly and dynamics. Cell, 185(24), 4560-4573.e19.
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2

Purification of Recombinant Human GRK5

Check if the same lab product or an alternative is used in the 5 most similar protocols
Full length wild-type human GRK5 was modified with a C-terminal hexa-histidine tag and cloned into pVL1392 vector for baculovirus production. GRK5 was expressed and purified as previously published46 . Briefly, Sf9 insect cells (Expression Systems) were infected with a BestBac-derived baculovirus at a density of 3.5 × 106 cells/mL and harvested 48 hours post infection. Cells were resuspended, lysed by sonication and the supernatant was applied to Ni-NTA resin. The resin was washed with lysis buffer and GRK5 eluted with lysis buffer supplemented with 200 mM imidazole. The combined eluate was then subjected to cation-exchange chromatography using a MonoS 10/100 column (GE healthcare) and eluted with a linear gradient of NaCl. Fractions containing GRK5 were combined and run on a Superdex 200 10/300 GL column (GE healthcare). GRK5 was aliquoted, flash frozen, and stored at −80 ˚C until use.
Huang W., Masureel M., Qianhui Q., Janetzko J., Inoue A., Kato H.E., Robertson M.J., Nguyen K.C., Glenn J.S., Skiniotis G, & Kobilka B.K. (2020). Structure of the Neurotensin Receptor 1 in complex with β-arrestin 1. Nature, 579(7798), 303-308.
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3

Purification of Recombinant Human GRK5

Check if the same lab product or an alternative is used in the 5 most similar protocols
Full length wild-type human GRK5 was modified with a C-terminal hexa-histidine tag and cloned into pVL1392 vector for baculovirus production. GRK5 was expressed and purified as previously published46 . Briefly, Sf9 insect cells (Expression Systems) were infected with a BestBac-derived baculovirus at a density of 3.5 × 106 cells/mL and harvested 48 hours post infection. Cells were resuspended, lysed by sonication and the supernatant was applied to Ni-NTA resin. The resin was washed with lysis buffer and GRK5 eluted with lysis buffer supplemented with 200 mM imidazole. The combined eluate was then subjected to cation-exchange chromatography using a MonoS 10/100 column (GE healthcare) and eluted with a linear gradient of NaCl. Fractions containing GRK5 were combined and run on a Superdex 200 10/300 GL column (GE healthcare). GRK5 was aliquoted, flash frozen, and stored at −80 ˚C until use.
Huang W., Masureel M., Qianhui Q., Janetzko J., Inoue A., Kato H.E., Robertson M.J., Nguyen K.C., Glenn J.S., Skiniotis G, & Kobilka B.K. (2020). Structure of the Neurotensin Receptor 1 in complex with β-arrestin 1. Nature, 579(7798), 303-308.
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