MCB 2050 Lecture Notes - Lecture 22: Sodium Dodecyl Sulfate, Western Blot, Interactome
21 Oct 2016
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Document Summary
Sodium dodecyl sulfate polyacrylamide gel electrophoresis (sds page) Protein interactome (iv) techniques for protein analysis. Cells are lysed in buffer containing the detergent sodium dodeycl sulfate. Sds binds proteins imparting a net negative charge, migrates to anode. Polypeptides migrate through a porous polyacrylamide gel matrix in an electric field. Mobility is a linear function of the logarithms of their molecular mass. Used to separate all polypeptides in a complex mixture. Protein bands in gels visualized by coommassie blue staining. Immunoblotting detection of a specific protein by specific antibody. Transfer of separated proteins from page to solid support (membrane) Detection of a target protein by a specific antibody. * the membrane creates an antibody for the protein. *how to detect the antibody that was made through the sds-page method. Protein gel sandwiched beside a blotting membrane. Negatively charged proteins migrate out of gel towards +ve electrode and trapped by blotting membrane.
