Question

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Draw the elution profile for a size exclusion column with an exclusion range of 50 kD to 500KD on which are separated. Make sure you indicate the bed and void volumes Myoglobin Hemoglobin Immunoglobulin Glutamine synthetase 17 kD -68 kD 150-200 kD 400 kD Draw the elution profile for an S sepahrose, cation exchange column pl 10.3 on which MW Cytochrome C Ribonuclease 12.0 kD 13.7 kD 7.8 9.6 14.7 kD Lysozyme Myoglobin 17.1 kD 7.5 Are separated in a pH=7 buffer 5.3 Chromatography Principles: Part 2 Gel Filtration Slide 78 of 217 Buffer Solvent Bed volume small proteins Gel Ge matrix bead Small 1) Exclusion Gel beads Volume of efffuent Large molecules limit 0 and larger, will elute together. 2) Lower limit ARR 5.3 Chromatography Principles: Part 2 Slide 86 of 217 Name Type Fractionation Range (kD) copy 2.docx unoun

Answer 1

Olusad DHCTAL HAAR ramge- 50-500KD uhorg Ad Erelusiom Oul500KD wag the exclusion 'mit Glutae looul m Ho emoplobin eric det o NA Junala 1 00 Frrantion Numlo es 00 Vid veume volume af he beads Ashich = Total volume beol voluume 00 Lobins noleeular weight the exelusion range loroer init thought Gel frtraton 7KD s 7.00- to pars Tthrogh ttbn coumn thue myoglobin pates all 8.00 the tho Absorbance

ber Monday Yu e Th Fr S so Tu W Th F s tHo T We Th Fr Sa SMo 12 ts 1s DECEMBER glustarni'ne Syedtaae (hook KD mobeular uoei quila a large Aronght qel amd so, thay wi emolacula 0amo to entel fevo poreo fesir Imunoglobulin 150-a00KDmo leeular weight AM the and witt elutad soones 9.00 medium Ahat uiu anle sorne pores n little later than moleeule 10.00 uril be euteol the and Riutabuine Sypatiase temoglobim(8KD) amolueular o 12.00 0eight proteil that enter amaxmidm oh andl so, usiu elulio nuno ayly wiu all lales 12.00- pores than n he gel Glustamine V Sythace tamulabulin iPM 2.00- of eolumn an aation case 3.00 Myoganite and Ripauelaase means at this pt have pI values7.5to子.8 Aid be qeutral orthey en them. So, elution Mil reouit n the elutien pich Noulorot have 4.00 any met a pt-7 with buffer ade 5,00 of glolin followed toy ibonuclease. Agein dyeoiyme (pT and Cytochrome e afher Gytoohre e d tham 6.00 wiu be wiu aftei the ne and Jbe eoCn anal 00 Y me the tound so, wil be more will be elulsol laler Ribonucrase - Cutoe yfnckroms e Fracthen Number