Protein Analysis Lab

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Suzanne Murray G00324885 Group D
Thursday 10 am lab 3/11/16
Introduction
The aim of this experiment is to separate the protein samples based on their molecular size using the SDS-PAGE technique and to detect EGFP protein by carrying out a western blot.
Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) is a technique used in the lab for the separation of proteins by their molecular weight. SDS is a detergent used in PAGE because its main role is to break down the disulphide bonds which disrupts the tertiary structure of the proteins making them into a linear shape. It then coats the protein with a negative charge which deactivates the charge that is present on the R groups of the amino acids. Once SDS has carried out their role the charge of the protein is then proportional to its molecular weight. (Oswald et al, 2016) …show more content…

It involves transferring or blotting g proteins that were separated using electrophoresis form the gel to a membrane where they can then be seen and examined. It is mainly used to identify if antibodies are present in a protein sample, which can then diagnose many conditions such as HIV and Lyme disease. It is mainly used in hospitals to do so. (The Human Protein Atlas, 2012)
The method used for staining in this lab is the CN/DAB development solution. It contains substrate and peroxide solutions for combined chloronaphthol and diaminobenzidine based detection of horseradish peroxidase (HRP) activity in blotting and tissue staining methods. This solution stains the protein bands a dark turquoise blue which makes them easy to

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