In immunoblotting, what are 3 reasons we first transfer the proteins from the ge
ID: 180316 • Letter: I
Question
In immunoblotting, what are 3 reasons we first transfer the proteins from the gel to a membrane? Two proteins ('C and 'D) differ slightly from one another. Protein 'C' is 10% larger in mass than Protein 'D while Protein 'D' has more positively-charged amino acid residues than Protein 'C'. The two proteins are isoforms of one another, meaning that structurally and functionally, they are very similar to one another. Design an isolation scheme that can be used to separate these two proteins from one another.Explanation / Answer
1. the mixture of proteins are transferred from a gel to a Nitrocellulose membrane for the identification of proteins in the given mixture.the protein mix transffered to nitrocellulose membrane are allowed to react with primary and secondary Antibodies,these antibodies reacted with specific proteins are then detected using staining with fluroscent stains.So Transfer is Important and essential for identification of proteins in the given sample.
2.Gel Electrophoresis can be employed to separate the proteins. gel electrophoresis can seperate the proteins based on size and charge, proteins are not negatively charged so we use sds(detergent) in seperating proteins.
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