Chemical & Enzymatic methods

B-per, BugBuster

detergent based reagents

High concentration detergent-based reagents

not compatible with protein assay and mass spectrometry in protein analysis

Lysozyme (200 ug

mL)

for the enzymatic method, we use _; a digest polysaccharide component of yeast and bacterial cell walls, making the cells easier to lyse

Glass beads

alternative method to enzymatic method especially when dealing with yeast cells

DNAse & RNAse

what is added to avoid interfering protein target when sample becomes viscous

Column chromatography

primary protein purification; combination of proteins and solutions added to a surface of a cylindrical column, loaded with a porous solid matrix that is submerged in a solvent

ion exchange chromatography

columns of ion exchange filled with tiny beads holding either (+) or (-) charges that retard protein of the opposite charge

gel filtration chromatography

separate proteins by size; the matrix are small porous beads

affinity chromatography

matrix is covalently paired with a molecule that interacts specifically with the protein of interest; proteins that attach specifically can be produced through a pH shift or conc. salt solutions; proteins are extremely purified