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5 × 5 ml º • HiTrap Capto adhere 5 × 1 ml º • 5 × 5 ml º • HiScreen Capto MMC 1 × 47 ml º HiScreen Capto adhere 1 × 47 ml º HiTrap Capto Core 700 5 × 1 ml º • HiScreen Capto Core 700 1 × 47 ml º Affinity chromatography • Recommended for system º Can technically be used with the system,Continued with the Superdex 75 (16/60) column having lower molecular mass range State of Aggregation and Glycosylation of IndiVidual Molecules Since the last step in all of the protein purification procedures involved gel filtration, there was no need to perform additional chromatography to determine the ag gregation states of the samples IL19 and IL wereFractions were collected and analysed by SDS–PAGE Fractions containing the IBD protein were concentrated to 2 ml using an Amicon Ultra15 concentration device with a 3 kDa molecularweight cutoff before the addition of a 01 1volume

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Superdex 75 16/60 pdf

Superdex 75 16/60 pdf-HiLoad 16/60 or 26/60 Superdex 75 prep grade HiLoad 16/60 or 26/60 Superdex 0 prep grade Dismantling tool, Support screens, net ringsμm), Oring, domed nuts Equilibration before a new run Regenerate the column after each run with one column volume of buffer at 30 cm/h (1 ml/min for XK 16/60 or 26 ml/min forXK 26/60) Buff ers and solvent resistance Degas and filter all solutionsGel filtration on a Superdex 75 HR 16/60 (Amersham Bio sciences) equilibrated with 01 M Tris/HCl, pH 80, 1 M KCl Pure protein fractions, as revealed by SDSPAGE and mass

Reference 29 1487 21 Superdex 75 Increase 10 300 Gl Dd Biolab Laboratory Equipment

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;Increased resolution compared with Superdex 75, forFind SigmaAldrichU MSDS, related peerreviewed papers, technical documents, similar products & more at SigmaAldrich

Flow rate needs to be decreased when working at low temperature or with viscous solutions, see product instructions for more details Table 3 Column choice for different applications Type of application/column 10/300 GL 5/150 GL 32/300 Smallscale preparative runs (mg) × MicroscaleSuperdex 0 Increase 5/150 GL 5 x 150 3 4 to 50 42 000 045 075 50 Superdex 0 Increase 32/300 32 x 300 24 4 to 50 48 000 0075 015 45 Table 3 Column choice for different applications Type of application/column 10/300 GL 5/150 GL 32/300 Smallscale preparative runs (mg) × Microscale preparative runs (µg) When sample amount is limited and small consumtionPowder should be dissolved in 6070% of the final volume, pH adjusted to 80, then water added to final volume Other stock solutions 02M 025M EDTA o The stock concentration is relatively low due to the low solubility of this compound Add 5M NaOH to the solution until all EDTA is dissolved (pH is 758) before you

Further purified by gel filtration using a 1ml of Superdex 75 16/60 prepgrade column (Amersham Pharmacia) on an AKTA purifier system (GE Healthcare), in 50mM Tris/HCl, pH 75, buffer containing 150mM NaCl and1mM EDTA Reduction and oxidation of PDI proteins PDI proteins were reduced with 10mM DTT for 10min at room temperature DTT wasHiLoad 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 Superose – Prepacked Tricorn and PC columns Superose 12 10/300 GL 1 × 103 to 3 × 105 24 100 Superose 12 PC 32/30 1 × 103 to 3 × 105 24HiLoad ® 16/600 Superdex ® 75 prep grade are prepacked 1 mL XK columns designed for highresolution preparative gel filtration chromatography separating proteins and polynucleotides Superdex ® prep grade (pg) is produced by covalent bonding of dextran to highly crosslinked agaroseThe separation properties of these media are determined by the dextran component

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60 min (04 mL/min) Superdex 30 Increase 10/300 GL min (12 mL/min) oeø Retention time (min) Columntocolumn Results from tests of column packing robustness are shown on Superdex 0 Increase 10/300 GL columns in the Figure Three columns from different lots, packed with the same resin lot, were tested and the results show very high reproducibility Flow rate OS mL/minHiLoad 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 crosslinked agarose The steep selectivity of dextran and the high chemical Column HiLoad 16/60 Superdex 75 prep grade Sample IGF1, ZZ fusion protein uncleavedmaterial Buffer 015 ammonium acetate, pH 60 Flow rate 075 ml/min (225 cm/h) Fig recombinantIGF1 (insulinlike growth factor 600)from its fusion protein partner (ZZ, daltons) uncleavedmaterial Columnvoid volume Totalcolumn volume 280nm 01 IGF1 10 Time ColumnHiLoad 16/60 Superdex 75

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Column a) HiLoad 16/60 Superdex 75 prep grade b) HiLoad 16/60 Superdex 0 prep grade Sample 1 Myoglobin 15 mg/ml Mw 17 000 2 Ovalbumin 4 mg/ml Mw 43 000 3 Albumin 5 mg/ml Mw 67 000 4 IgG 02 mg/ml Mw 158 000 5 Ferritin 024 mg/ml Mw 440 000 Sample vol 05 ml Buffer 015 M NaCl, 005 M phosphate buffer pH 70 001% sodium azide Flow rate 15 ml/min 10 12 14 16 18 40 60Selectivity curves from Superdex 30 prep grade, Superdex 75 prep grade, and Superdex 0 prep grade Fig 4 Separation of test substances on HiLoad 26/600 Superdex 30 pg Superdex 30 prep grade resin is optimized for proteins/peptides below M r 10 000 Fig 5 Comparison between the selectivity of Superdex 75 prep grade and Superdex 0 prepThis Amersham (GE Healthcare) SuperDex 75 Prep Column is new from surplus stock It ships in the original packaging It comes with the instruction sheet and XK16 Accessory Kit () The column is labeled XK 16 HiLoad 16/60 Superdex 75 Prep Grade Part Number More information can be found on the Amersham/GE website

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Hi Load Superdex 75 16 60 Prep Grade Size Exclusion Column, supplied by GE Healthcare, used in various techniques Bioz Stars score 99/100, based on 1 PubMed citations ZERO BIAS scores, article reviews, protocol conditions and moreNPC1C 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 full yglycosylated NPC1C protein wasSuperdex 75 Increase prepacked columns are designed for rapid separation for smallscale preparative purification and analysis of recombinant tagged or other proteins with molecular weights (Mr) from 3000 up to 70,000 Comprising smaller, more rigid beads with a narrower particle size distribution and a higher selectivity;

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 The eluted protein was pooled, concentrated and loaded on a Superdex 75 16/60 column (GE Healthcare) equilibrated with buffer C (50 Article Download PDF View Record in Scopus Google Scholar 2 A Israël The IKK complex, a central regulator of NF κB activation Cold Spring Harb Perspect Biol, 2 (10), p a CrossRef View Record in Scopus GoogleQuick information 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 (Superdex 75 pg, HiLoad 16/60 Superdex 0 pg, and MonoQ HR 5/5 were from GE Healthcare Restriction endonucleases and PCRrelated enzymes were from Toyobo Amicon Ultra4 centrifugal filters were from Millipore BioGel P2 was from BioRad Laboratories Other analytical grade chemicals were obtained from commercial sources Microorganisms and Culture Conditions–

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GE Healthcare hiload 16 60 superdex 75 column Hiload 16 60 Superdex 75 Column, supplied by GE Healthcare, used in various techniques Bioz Stars score 86/100, based on 1 PubMed citationsSuperdex 75 16/60 a 10/300 GLor column (GE Healthcare) in mM Tris pH 75 containing 0 mM NaCl and 1 mM DTTSOD1 eluted from columnboth s as a single peak (~115 mL corresponding to ~32 kDa, ie dimer, on 10/300 GL columnthe and the fractions containing ) pure protein (≥ 90 %), as judged by SDSPAGE, were combined Matrix assisted laser desorption ionization timeofAntibody Purification Handbook The Recombinant Protein Handbook Protein Amplification and Simple Purification Protein Purification Handbook Ion Exchange Chromatography Principles and Methods Affinity Chromatography Principles and Methods Hydrophobic Interaction Chromatography

Figure 7 3 From Size Exclusion Chromatography Semantic Scholar

Reference 29 1487 21 Superdex 75 Increase 10 300 Gl Dd Biolab Laboratory Equipment

Superdex® Gel Filtration Column phase HiLoad 16/60 Superdex 75 PG, L × ID 60 cm × 16 mm, 35 μm particle size;Ver 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 49Superdex 75 Increase 32/300 32 × 300 24 > 43 000 4 to 50 0075 015 * Note!

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 We have HiLoad Superdex 0 16/60 Gel Filtration column in our lab, which have some 'Sticky Proteins' and protein aggregates bound to the columnHiLoad 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 crosslinked agarose The steep selectivity of dextran and theCan be used with standalone pump or chromatography system High resolution – for high purity Scalable – separations can be scaled up using Superdex 75 prep grade resin

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000 Base CV 1637 {ml} HiLoad_16/60_Superdex_75_prep_grade 000 Block Inject_Peak (Inject_Peak) 000 Base Volume 000 InjectionValve Reinject 000 LoopSelection LP1 000 Set_Mark "Peak_injection" 1000 InjectionValve Inject 1000 End_Block 030 Block Fractionation_Gel_Filtration (Fractionation_Gel_Filtration) 000 Base CV 1637 {ml} HiLoad_16/60_Superdex_75Injection onto a HiLoad Superdex 75 pg 16/60 column (GE Healthcare, UK) and elution in mM Tris pH 75, 0 mM NaCl, 1 mM TCEP;HiLoad 16/600 and 26/600 Superdex 30, 75, and 0 prep grade Instruction for Use 2917 AF 3 CleaningInPlace (CIP) Regular cleaning Wash the column with onehalf to one CV 05 M NaOH at a flow rate of 08 mL/min for 16/600 or 22 mL/min for 26/600 to remove most of the nonspecifically bound proteins from the chromatography resin After cleaning, immediately

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