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200 10 300 gl column

Manufactured by GE Healthcare
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The 200 10/300 GL column is a chromatography column designed for size exclusion chromatography. It has a bed volume of 300 ml and an internal diameter of 10 mm. The column is constructed with a glass column body and is compatible with aqueous and organic solvents.

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Lab products found in correlation

Rdr2 and protein standards, by GE Healthcare (2 mentions) Ktamikro, by GE Healthcare (1 mentions) Ktaprime, by GE Healthcare (1 mentions)

4 protocols using 200 10 300 gl column

1

MutL Protein Oligomerization Assay

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MutL WT and MutLΔ4 (1.5 mg/ml) in 25 mM HEPES–KOH [pH 7.5], 150 mM KCl, 5 mM MgCl2, 10% glycerol and 10 mM β-mercaptoethanol (GF buffer) were mixed with 1 mM adenylylimidodiphosphate (AMPPNP) or GF buffer as control and incubated for 16 h at 4°C. Samples (50 μl) were injected on a Superdex 200 10/300 GL column equilibrated with GF buffer and operated by ÄktaMikro (GE Healthcare) at 50 μl/min. Elution was monitored at 280 nm.
Mardenborough Y.S., Nitsenko K., Laffeber C., Duboc C., Sahin E., Quessada-Vial A., Winterwerp H.H., Sixma T.K., Kanaar R., Friedhoff P., Strick T.R, & Lebbink J.H. (2019). The unstructured linker arms of MutL enable GATC site incision beyond roadblocks during initiation of DNA mismatch repair. Nucleic Acids Research, 47(22), 11667-11680.
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2

Molecular Mass Estimation by FPLC

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To estimate RDR2’s mass in solution, RDR2 and protein standards (GE Healthcare) were subjected to FPLC using a Superdex 200 10/300 GL column at 4 °C using 50 mM Hepes-KOH pH 7.5, 150 mM NaCl as the running buffer and a flow rate of 0.3 mL/min. Protein peaks were detected by UV absorbance at 280 nm, and SDS-PAGE with staining by Coomassie Blue.
Mishra V., Singh J., Wang F., Zhang Y., Fukudome A., Trinidad J.C., Takagi Y, & Pikaard C.S. (2021). Assembly of a dsRNA synthesizing complex: RNA-DEPENDENT RNA POLYMERASE 2 contacts the largest subunit of NUCLEAR RNA POLYMERASE IV. Proceedings of the National Academy of Sciences of the United States of America, 118(13), e2019276118.
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3

Purification via Size Exclusion Chromatography

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Size exclusion chromatography was performed on a GE ÄKTAprime plus FPLC system applying with a Superdex 200 10/300 GL column at a 0.5 mL/min flow rate. Detection was at 280 nm and 0.25 mL fractions were collected.
Swiecicki J.M., Santana J.T, & Imperiali B. (2019). A Strategic Approach for Fluorescence Imaging of Membrane Proteins in a Native-Like Environment. Cell chemical biology, 27(2), 245-251.e3.
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4

FPLC-Based Mass Estimation of RDR2

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To estimate RDR2's mass in solution, RDR2 and protein standards (GE Healthcare) were subjected to FPLC using a Superdex 200 10/300 GL column at 4˚C using 50 mM HEPES-KOH pH7.5, 150 mM NaCl as the running buffer and a flow rate of 0.3 ml/min). Protein peaks were detected by UV absorbance at 280 nm, and SDS-PAGE with staining by Coomassie Blue.
Mishra V., Singh J., Fukudome A., Wang F., Zhang Y., Takagi Y., Trinidad J.C., & Pikaard C.S. (2020). Assembly of a double-stranded RNA synthesizing complex: RNA-DEPENDENT RNA POLYMERASE 2 docks with NUCLEAR RNA POLYMERASE IV at the clamp domain.
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