Experimental Procedures Of An Ige Antibody

2190 Words May 17th, 2015 9 Pages
There were several experimental procedures that were performed by Bayry et al that allowed for this experiment to be performed. One of these procedures included treatment of an IgE antibody that is produced specifically in mice, SPE7 with heme using the technique immunoblotting to determine the polyreactivity of heme and the resulting consequences of the reaction. Immunoblotting, also known as western blotting, is a type of an assay that is specifically used for the detection and characterization of proteins1. According to Gallagher and Chakavarti, immunoblotting works by exploiting the specificity inherent in antigen-antibody recognition5. It involves the solubilization and electrophoretic separation of proteins, glycoproteins, or lipopolysaccharides by gel electrophoresis, followed by quantitative transfer and irreversible binding to nitrocellulose, PVDF, or nylon5. The immunoblotting technique has been useful in identifying specific antigens recognized by polyclonal or monoclonal antibodies and is highly sensitive5. This unit provides protocols for protein separation, blotting proteins onto membranes, immunoprobing, and visualization5. Another technique used was an Enzyme-Linked Immunosorbent Assay or ELISA. The purpose of an ELISA is to determine if a particular protein is present in a sample and if so, how much6. There are two main variations on this method and whichever one is used depends on the type of experiment that is being run. One can determine how much…
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