BIOL 2083 Lecture Notes - Lecture 3: Equilibrium Constant, Leucine, Acrylamide
9 Oct 2014
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Determinations of: molecular weight charge primary sequence disulfides. Using the relative densities of different cellular fractions. Cellulose acetate dialysis bags with controlled pore sizes (mwco) Used qualitatively for removing small molecules from proteins (salts, ligands) Can be used quantitatively to study the equilibrium constant for binding of small molecules to proteins. Cellulose beads with controlled pore sizes are poured into a column. Proteins and other molecules can enter the beads if they are smaller than the pore size. Distance they must travel through the column is longer. Cellulose (or other matrix) beads with attached charge groups (amino or carboxyl groups) Attaching a molecule to the matrix that specifically and uniquely interacts with your protein. The protein can be eluted using a solution of the the same molecule that is attached to the beads, or by changing the solution conditions to ones that do not favor binding. Elution from any of these columns is generally monitored by spectroscopic methods.
