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Today: Lab notebooks due. Next week: M/T analyze gels Computer lab Turn in a one paragraph summary of your denaturation experiment (per PAIR) W/R CD: each PAIR will sign up for 25 minutes of CD time. Figure 6-11 Use of dialysis to separate small and large molecules. Page 139.
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Today: Lab notebooks due Next week: M/T analyze gels Computer lab Turn in a one paragraph summary of your denaturation experiment (per PAIR) W/R CD: each PAIR will sign up for 25 minutes of CD time
Figure 6-11 Use of dialysis to separate small and large molecules. Page 139
Figure 6-29 A selection of preparative ultracentrifuge rotors. Page 154
Figure 6-32 Isopycnic ultracentrifugation. Page 155 components of a mixture are separated on the basis of differences in density
Figure 6-28 Sedimentation coefficients in Svedbergs (S) for some biological materials. Page 153
Theoretical considerations http://www.bergen.org/AAST/Projects/Gel/technique2.htm
Ammonium persulfate: initiates the polymerization NH4SO3-O-O-SO3NH4 S2O8-2 + 1e- SO4-2 + SO4-• Free radical TEMED: N,N-tetramethylenediamnine: catalyst (CH3)2- N-CH2-CH2-N(CH3)2 • SO4-• + n CH2=CH-CONH2 X-CH2-CH-CONH2 • X-CH2-CH(CONH2)-CH2-CH(CONH2)-CH-CONH2
Figure 6-20 Apparatus for slab gel electrophoresis. Page 146
Structural features of most globular proteins: 1. Very compact: e.g. Mb has room for only4 water molecules in its interior. 2. Most polar/charged R groups are on the surface and are hydrated. 3. Nearly all the hydrophobic R groups are on the interior. 4. Pro occurs at bends/loops/random structures and in sheets
Simulation http://www.rit.edu/~pac8612/electro/Electro_Sim.html
Figure 6-25 Logarithmic relationship between the molecular mass of a protein and its relative electrophoretic mobility in SDS-PAGE. log
Proteins in native gels exhibit a more complex relationship between mobility and MW.
Figure 6-27 Two-dimensional (2D) gel electrophoresis. Page 150