2.2EI Clectrophoresis separation is nearl always carried out in gels(rather than in free solutions) 2.2.1 The chemically inert polyacrylamide gel, formed by the polymerization of acrylamide monomer cross linked by methylenebisacrylamide, is routinely used 2.2.2 The pore size of polyacrylamide gel can be controlled by choosing various concentrations of acrylamide and methylenebisacrylamide at the time of polymerization(less cross-linking agent, methylenebisacrylamide, larger pore size)
2.2 Electrophoresis separation is nearly always carried out in gels (rather than in free solutions). 2.2.1 The chemically inert polyacrylamide gel, formed by the polymerization of acrylamide monomer cross linked by methylenebisacrylamide, is routinely used. 2.2.2 The pore size of polyacrylamide gel can be controlled by choosing various concentrations of acrylamide and methylenebisacrylamide at the time of polymerization (less cross-linking agent, methylenebisacrylamide, larger pore size)
2.2.3 Gels suppress convective currents produced by small temperature gradients, a requirement for effective separation Gels also prevent diffusion 2. 2. 4 Gels serve as molecular sieves that enhance separation. 2.2.5 Arne Tiselius won the nobel prize in Chemistry in 1948 for inventing the electrophoresis technique and applying it to protein studies
2.2.3 Gels suppress convective currents produced by small temperature gradients, a requirement for effective separation. Gels also prevent diffusion. 2.2.4 Gels serve as molecular sieves that enhance separation. 2.2.5 Arne Tiselius won the Nobel Prize in Chemistry in 1948 for inventing the electrophoresis technique and applying it to protein studies
2.3 SDS-polyacrylamide gel electrophoresis (PAGE is commonly used for estimation of protein purity and molecular mass. 2.3.1 The mixture of proteins is first dissolved in a solution of sDs (sodium dodecyl sulfate an anionic detergent that disrupts nearly all noncovalent interactions in native proteins
2.3 SDS-polyacrylamide gel electrophoresis (PAGE) is commonly used for estimation of protein purity and molecular mass. 2.3.1 The mixture of proteins is first dissolved in a solution of SDS (sodium dodecyl sulfate), an anionic detergent that disrupts nearly all noncovalent interactions in native proteins
2.3.2 Mercaptoethanol or dithiothreitol is also added to reduce disulfide bonds 2.3.3 Anions of sds bind to peptide main chains at a ratio of a bout one sds for every two amino acid residues, which gives a complex of sds with a denatured protein a large net negative charge that is roughly proportional to the mass of the protein
2.3.2 Mercaptoethanol or dithiothreitol is also added to reduce disulfide bonds. 2.3.3 Anions of SDS bind to peptide main chains at a ratio of about one SDS for every two amino acid residues, which gives a complex of SDS with a denatured protein a large net negative charge that is roughly proportional to the mass of the protein
O Na o-s0-(CH2)1 CH3 Sodium dodecyl sulfate (SDS)