MCB 102 F08 Lect _5 DRAFT

MCB 102 F08 Lect _5 DRAFT - Announcements None at the...

Info iconThis preview shows pages 1–7. Sign up to view the full content.

View Full Document Right Arrow Icon
Announcements None at the moment, as of 8 PM, Sunday, 7 Sept. 2008
Background image of page 1

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
Procedures for Sequential Fractionation of Proteins •Enrichment of appropriate subcellular fraction (centrifugation) •Appropriate extraction conditions (solubilization in buffer) •Dialysis (removal of small molecules) •Salting out (precipitation using ammonium sulfate) •Ion exchange chromatography (anion and cation) •Affinity chromatography (specific ligand binding) •Size exclusion chromatography (Stokes radius)
Background image of page 2
3,000 35 30,000 52,500 1.75 affinity column eluate 110 50 1,100 75,000 68.8 size exclusion eluate 9 77 90 115,500 1,278 ion exchange eluate 3 92 30 138,000 4,600 (NH 4 ) 2 SO 4 precipitate 1 100 10 150,000 15,000 Homogenate Purification (-fold) Yield (%) Specific Activity (units/mg) Total Activity (units) Total Protein (mg) Step Tracking the Purification Protocol of a Fictitious Protein
Background image of page 3

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
(— —) [KCl] (M) (- - -) (——)
Background image of page 4
Determination of Protein Concentration from UV Absorbance The Beer-Lambert Law: Absorbance ( A ) = log I 0 /I =  ε cl where ε = molar extinction coefficient of the protein (usually at 280 nm) c = concentration of the protein ( M ) l = path length of the cuvette (usually 1 cm)  
Background image of page 5

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
Practically, however, especially at the early stages of
Background image of page 6
Image of page 7
This is the end of the preview. Sign up to access the rest of the document.

This document was uploaded on 10/11/2008.

Page1 / 23

MCB 102 F08 Lect _5 DRAFT - Announcements None at the...

This preview shows document pages 1 - 7. Sign up to view the full document.

View Full Document Right Arrow Icon
Ask a homework question - tutors are online