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protein purification and antimicrobial assay
Posted by: zia (IP Hidden, Unregistered user, )
Date: March 15, 2005 10:50AM
respected sir,
i am working on very small amino acids sequencing protein (antifungal protein), but the problem is elution and purification of proteins. after centrifugation these proteins are present in supernatent and the next step is presipitation of proteins. what method is best either by salt or organic solvent. the problem is that the required protein is present in very small amount and in supernatent a large amount of different protein is present. what i do in this case? electrophoresis is good method for this or hplc. but the problem in hplc is that there is no specific methedology as of mobile phase or elution time and so on. so i think electrophoresis is best. Second kindlt tell me the best method for antimicrobial method for protein antimicrobial (antibacterial and antifungal) assay, eith disc slip, gel diffusion or what? comment please zia
Re: protein purification and antimicrobial assay
Posted by: bassamfahmawi (IP Hidden, Regular member, 42)
Date: March 15, 2005 08:14PM
Hi,
Use the antibacterial minimal inhibitory concentartion method emplyed for testing antibiotic sensitivity and resistance on gram postive and Negative bacteria. In your situation, you can first detrmine the concetration your protein concantration and soak Whatman filter paper Mini discs with your protein preparatio and test them on Muller Hinton media against E.coli, Staph aurues etc.., exactly similar to antibiotic snsitivity assay. Check the latest edition of the ASM clinical microbiology Manual. Good Luck!
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