1) Draw a chromatogram depicting separation of proteins 1, 2, and 3 (protein #1, mw 30,000, protein #2, mw 60,000, and protein #3, mw 90,000). b)Indicate on the chromatogram in (a) where insulin would elute. Would insulin have total accessibility, some accessibility or no accessibility to the particle pores on this column? c)Referring to the size exclusion chromatogram in (a), indicate on the chromatogram where you would expect a molecule with a size of 40,000 to elute. Would you expect the molecule to be well resolved, somewhat resolved, or not resolved from the peak for protein #1?

Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
Section: Chapter Questions
Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
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1) Draw a chromatogram depicting separation of proteins 1, 2, and 3

(protein #1, mw 30,000, protein #2, mw 60,000, and protein #3, mw 90,000).

b)Indicate on the chromatogram in (a) where insulin would elute. Would insulin have total accessibility, some accessibility or no accessibility to the particle pores on this column?

c)Referring to the size exclusion chromatogram in (a), indicate on the chromatogram where you would expect a molecule with a size of 40,000 to elute. Would you expect the molecule to be well resolved, somewhat resolved, or not resolved from the peak for protein #1?

TABLE 26-4 Representative molecular exclusion media
Gel filtration in open columns
TSKgel SW silica for HPLC
Fractionation range for
Fractionation range for
globular proteins (Da)
5 000-150 000
10 000-500 000
Pore size
Name
globular proteins (Da)
Name
(nm)
Sephadex G-10
Sephadex G-25
Sephadex G-50
Sephadex G-75
Sephadex G-100
Sephadex G-200
to 700
1 000-5 000
1 500-30 000
G2000SW
G3000SW
12.5
25
45
G4000SW
20 000-7 000 000
3 000-80 000
4 000-150 000
5 000-600 000
NOTE: Sephadex is manufactured by GE Amersham Biosciences. TSKgel SW silica is manufactared by Tosoh Bioscience.
Transcribed Image Text:TABLE 26-4 Representative molecular exclusion media Gel filtration in open columns TSKgel SW silica for HPLC Fractionation range for Fractionation range for globular proteins (Da) 5 000-150 000 10 000-500 000 Pore size Name globular proteins (Da) Name (nm) Sephadex G-10 Sephadex G-25 Sephadex G-50 Sephadex G-75 Sephadex G-100 Sephadex G-200 to 700 1 000-5 000 1 500-30 000 G2000SW G3000SW 12.5 25 45 G4000SW 20 000-7 000 000 3 000-80 000 4 000-150 000 5 000-600 000 NOTE: Sephadex is manufactured by GE Amersham Biosciences. TSKgel SW silica is manufactared by Tosoh Bioscience.
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