The oligomeric state of rPfen was analyzed by gelfiltration chromatography on a Superdex75 Hiload16/60 column on an AmershamPharmacia Biotech (Kwai Chung, Hong Kong), AKTA FPLC system The column was preequilibrated with 2 column vols buffer (50 m m sodium phosphate, 150 m m NaCl, pH 74)Superdex 75 Increase 32/300 32 × 300 24 > 43 000 4 to 50 0075 015 * Note!Cell Host & Microbe, Volume 17 Supplemental Information Structural Conservation Despite Huge Sequence Diversity Allows EPCR Binding by the PfEMP1
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Superdex 75 16/60 manual
Superdex 75 16/60 manual-Superdex 75 10/300 GL and Superdex 0 10/300 GL are Tricorn™ high performance columns The columns are prepacked glass columns for high performance gel filtration of proteins, peptides, DNA fragments ( The denatured protein was purified on cobalt affinity resin (Clontech Laboratories) and refolded via dialysis in lysis buffer plus 5 mM DTT The Histag was removed by cleavage with TEV protease and the protein were further purified by gelfiltration chromatography (Superdex 75 HiLoad 16/60, GE Healthcare)
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HiLoad 16/600 and 26/600 Superdex 30 prep grade, HiLoad 16/600 and 26/600 Superdex 75 prep grade, and HiLoad 16/600 and 26/600 Superdex 0 prep grade (pg) are prepacked XK columns designed for preparative gel filtration Superdex prep grade is a composite matr ix of dextran and highly crosslinkedSuperdex gel filtration media are listed in Appendix C, Table 1 With their high physical and chemical stability, very high batchtobatch reproducibility, and Regulatory Support File backup, Superdex 30 prep grade, Superdex 75 prep grade and Superdex 0 prep grade are ideal for all stages of an industrial scale operation from research and The excess imidazole was removed by overnight dialysis and the sample was subjected to secondary subtractive NiNTA affinity chromatography step to remove the protease and uncleaved protein Finally, the protein was subjected to a gel filtration step using Superdex 75 16/60 column in a buffer containing 10 mM TrisHCl at pH 8, 150 mM NaCl
Superdex 75 Increase 10/300 GL is suitable for small scale preparative purification (μgmg) as a final polishing step, as well as for protein analysis and characterization The columns are supplied with two fingertight connectors 1/16" male for connection to ÄKTA ™ or other systems Table 1Resin data Matrix Composite of crosslinkedHiLoad 16/600 and 26/600 Superdex 30 prep grade, HiLoad 16/600 and 26/600 Superdex 75 prep grade, and HiLoad 16/600 and 26/600 Superdex 0 prep grade (pg) are prepacked XK columns designed for preparative size exclusion chromatography Superdex prep grade is a composite matrix of dextran and crosslinkedProduct Support HiLoad Superdex 75 pg prepacked columns are for highresolution size exclusion chromatography of recombinant proteins from samples up to 5 mL or 13 mL Prepacked format – for convenience and reproducibility;
Cm/h (1 ml/min for 16/60 or 26 ml/min for 26/60) Introduction HiPrep™ 16/60 and 26/60 Sephacryl™ S100 High Resolution (HR), Sephacryl S0 HR, Sephacryl S300 HR, Sephacryl S400 HR and Sephacryl S500 HR are prepacked gel filtration columns designed for preparative purification of peptides, proteins and larger moleculesSuperdex 75 Increase 32/300 1 Add to Cart Superdex 75 Increase 10/300 GL 1 Add to Cart 2333 HiLoad 16/600 Superdex 75 pg 1 × 1 ml Add to Cart HiPrep 16/60 Sephacryl S0 HR 1 ml Add to Cart HiPrep 16/60 Sephacryl S100 HR 1 ml Add to Cart 2334 HiLoad 26/600 Superdex 75 pg 1 × 3 ml Add to CartSuperdex 0 PC 32/30 1 × 104 to 6 × 105 24 10 Superdex prep grade – HiLoad™ prepacked columns HiLoad 16/60 Superdex 75 pg 3 × 103 to 7 × 104 1 500 HiLoad 26/60 Superdex 75 pg 3 × 103 to 7 × 104 3 1000 HiLoad 16/60 Superdex 0 pg 1 × 104 to 6 × 105 1 500 HiLoad 26/60 Superdex 0 pg 1 × 104 to 6 × 105 3 1000
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Refolding Of Proteins Using Gel filtration
Superdex ® 75 prep grade gel filtration medium offers very high resolution fractionation for separation, polishing and product formulation of recombinant proteins Superdex ® prep grade is a preparative gel filtration medium with a composite matrix of dextran and agarose This matrix combines the excellent gel filtration properties of crosslinked dextran (Sephadex) with theSuperdex 75 Increase 32/300, Superdex 75 Increase 5/150 GL, and Superdex 75 Increase 10/300 GL columns Selectivity of the resin Superdex 75 Increase and the related Superdex 0 Increase and Superose™ 6 Increase belong to the new generation of SEC resins based on highflow agarose with small beads TheFluorescent protein (FP)based biosensors based on the principle of intramolecular Förster resonance energy transfer (FRET) enable the visualization of a variety of biochemical events in living cells The construction of these biosensors requires
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NPC1C protein was then concentrated and purified by gel filtration on a HiLoad 16/60 Superdex ® 75 PG column (GE Healthcare) The NPC1C mutants were constructed by sitedirected mutagenesis kit (New England Biolabs), and then were expressed, refolded and purified following the procedures of wildtype NPC1C protein The fullSuperdex 30 pg up to ~ 10 000 Superdex 75 pg ~ 500 to 30 000 ~ 3000 to 70 000 Superdex 0 pg ~ 1000 to 100 000 ~ 10 000 to 600 000 Particle size, d50V1 ~ 34 µm Matrix Crosslinked agarose, spherical Pressure/ flow characteristics 40 to 60 cm/h atFind SigmaAldrichU MSDS, related peerreviewed papers, technical documents, similar products & more at SigmaAldrich
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Purification And Interactions Of The Muca And Mucb Proteins Constituting The Dna Polymerase Ri Genes And Environment Full Text
and further purified by gel filtration (HiLoad 16∕60 Superdex 75 column, GE Life Sciences) chromatography in buffer A ( mM Tris, 0 mM NaCl, 5 mM DTT, pH 80) The protein was concentrated to 10 mg∕mL The purity of the protein was >98% judged by SDSPAGE gel Suicide Probe Expression and Purification pTYB2 and pTXB1 The fraction containing the protein was then concentrated before injecting into a Superdex 75 16/60 sizeexclusion chromatography column preequilibrated with 10 mM HEPES pH 75, 150 mM NaCl, 1 mMVer A (LIT102) US/EG 0517 Sig 1216 Web site bioradcom USA 1 800 424 6723 Australia 61 2 9914 2800 Austria 43 1877 01 177 Belgium 32 (0)3 710 53 00 Brazil 55 11 3065 7550 Canada 1 905 364 3435 China 86 21 6169 8500 Czech Republic 4 241 430 532 Denmark 45 44 52 10 00 Finland 358 09 804 22 00 France 33 01 47 95 69 65 Germany 49
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% ethanol (for HiLoad Superdex 30 pg and Superdex 75 pg, use 0 mM sodium acetate in % ethanol) Note Use a lower flow rate for viscous % ethanol Store the column in % ethanol to prevent any microbial growth Connect the transport tool to the column outlet, to prevent air entering the column Fill the transport tool up toFlow rate needs to be decreased when working at low temperature or with viscous solutions, see product instructions for more details HiLoad 26/60 Superdex 75 prep grade HiLoad 16/60 Superdex 0 prep grade HiLoad 26/60 Superdex 0 prep grade Superdex™ 75 10/300 GL Superdex
Engineered Synthetic Antibodies As Probes To Quantify The Energetic Contributions Of Ligand Binding To Conformational Changes In Proteins Journal Of Biological Chemistry
Superdex 75 10 300 Gl And 5 150 Gl Cytiva
