Survey
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
Download Gel Electrophoresis - PHS International Baccalaureate
Document related concepts
Transcript
Gel Electrophoresis Pensacola High School IB Chemistry 3 Steps • Breakdown the protein into amino acids – Break amide/peptide bond – Called hydrolysis (opposite of condensation) – Acid or base and heat required • Add protein to gel containing a buffer • Apply a voltage – Amino acids move based on mass and charge • Dye amino acids – Ninhydrin • Measure distance traveled, • Compare to known Set-up Why amino acids move • Amino acids separate based on their isoelectric point and molar mass • Isoelectric point: – This is the pH where they net charge of amine and carboxylic acid groups cancel out +NH3-C-COO- • pH of buffer is more basic than isoelectric point, than amino acid will have a negative charge and move toward postive electrode – There are fewer hydrogen atoms around to protonate it NH2-C-COO- • pH of buffer is more acidic than isoelectric point, than amino acid will have a positive charge and move to negative electrode – More H atoms around to protonate +NH3-C-COOH
