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Fplc superose 6 column

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The FPLC Superose 6 column is a size exclusion chromatography column designed for the separation and purification of proteins, peptides, and other biomolecules. The column features a porous agarose-based matrix that allows for efficient separation based on the size and molecular weight of the analytes. The Superose 6 column is compatible with fast protein liquid chromatography (FPLC) systems and can be used with a variety of aqueous mobile phases.

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Lab products found in correlation

Centricon 10 000 kda cutoff, by Merck Group (2 mentions) Ab7970, by Abcam (2 mentions) Proteinase inhibitor cocktail, by Roche (2 mentions) Colorimetric assay, by Abcam (1 mentions) Uv detector, by Waters Corporation (1 mentions) 0.2 μm membrane, by Sartorius (1 mentions)

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Superose 6 column (200 citations) Superose 6 (82 citations) FPLC columns (9 citations)

4 protocols using fplc superose 6 column

1

Analysis of FoxP3 Interactions

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Activated CD4+ T cells transduced with FoxP3 or mutants (1×109 cells/sample) were lysed in NP-40 lysis butter (20mM Tris-HCl, 2mM EDTA, 1% NP-40 10% glycerol, 150mM NaCl, 1mM PMSF and 1X Proteinase inhibitor cocktail; Roche, #04693116001) for 2 hr with gentle vortexing at 4°C. The lysates were sedimented at 15,000 rpm for 30 min at 4°C to remove aggregates, and the supernatant injected into an FPLC Superose 6 column (10 X 300mm, GE Healthcare Life Science) at a flow rate of 400ul/min in PBS. Twenty seven fractions of 400 ul were collected, immediately concentrated with filter centrifugation (Centricon 10,000 KDa cutoff, Millipore) down to 50ul, resolved by SDS-PAGE and probed by sequential immunoblotting for FoxP3 (eBioscience, #14-5773). FoxP3 containing fractions were incubated with anti-RELA (Abcam, #ab7970) or anti-EZH2 (Active motif, #AC22), revolved by SDS-PAGE and immunoblotted with anti-FoxP3.
Kwon H.K., Chen H.M., Mathis D, & Benoist C. (2017). Different molecular complexes that mediate transcriptional induction and repression by FoxP3. Nature immunology, 18(11), 1238-1248.
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2

Analysis of FoxP3 Interactions

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Activated CD4+ T cells transduced with FoxP3 or mutants (1×109 cells/sample) were lysed in NP-40 lysis butter (20mM Tris-HCl, 2mM EDTA, 1% NP-40 10% glycerol, 150mM NaCl, 1mM PMSF and 1X Proteinase inhibitor cocktail; Roche, #04693116001) for 2 hr with gentle vortexing at 4°C. The lysates were sedimented at 15,000 rpm for 30 min at 4°C to remove aggregates, and the supernatant injected into an FPLC Superose 6 column (10 X 300mm, GE Healthcare Life Science) at a flow rate of 400ul/min in PBS. Twenty seven fractions of 400 ul were collected, immediately concentrated with filter centrifugation (Centricon 10,000 KDa cutoff, Millipore) down to 50ul, resolved by SDS-PAGE and probed by sequential immunoblotting for FoxP3 (eBioscience, #14-5773). FoxP3 containing fractions were incubated with anti-RELA (Abcam, #ab7970) or anti-EZH2 (Active motif, #AC22), revolved by SDS-PAGE and immunoblotted with anti-FoxP3.
Kwon H.K., Chen H.M., Mathis D, & Benoist C. (2017). Different molecular complexes that mediate transcriptional induction and repression by FoxP3. Nature immunology, 18(11), 1238-1248.
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3

Plasma Lipid and Lipoprotein Analysis

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Total cholesterol levels in plasma were measured using a colorimetric assay (BioVision). Plasma triglycerides (TG) were measured enzymatically according to the manufacturer’s instruction (Pointe Scientific). To determine total apolipoprotein distributions as an indication of changes in lipoprotein profiles, pooled plasma (n=4) was separated by a Superose 6 FPLC column (10/300 GL, GE Healthcare Life Sciences; flow rate 0.15 mL/min) coupled with a UV detector (Waters) for protein absorbance at 280 nm. For TG determination in the liver, total lipid was extracted from ~100 mg of tissue (n=8/group) in chloroform: methanol (2:1) solution and processed as described previously [22 (link)].
Fan R., Kim J., You M., Giraud D., Toney A.M., Shin S.H., Kim S.Y., Borkowski K., Newman J.W, & Chung S. (2019). α-linolenic acid-enriched butter attenuated high fat diet-induced insulin resistance and inflammation by promoting bioconversion of n-3 PUFA and subsequent oxylipin formation. The Journal of nutritional biochemistry, 76, 108285.
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4

Gel Filtration for Brain Protein Extraction

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Gel filtration was performed as previously described.77 (link) In brief, brain extracts were filtered through a 0.2-μm membrane (Sartorius, Goettingen, Germany). Gel filtration was carried out using a Superose 6 FPLC column (AKTA; GE Healthcare), and 0.25-ml fractions were collected.33 (link)
Jeong E.I., Chung H.W., Lee W.J., Kim S.H., Kim H., Choi S.G, & Jung Y.K. (2016). E2-25K SUMOylation inhibits proteasome for cell death during cerebral ischemia/reperfusion. Cell Death & Disease, 7(12), e2573-.
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