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I have a question about the recombinant human SSAT protein. On the reference page, it has a good picture of Western Blot which shows the protein around 25KD. I would like to know the details of the Western Blot, such as the detailed ingredients of the gel, running buffer, loading buffer, and the exact conditions as well. I was trying to use this as a positive control for my study. But every time I only got a very very faint band with 100ng of SSAT. Wre you able to give us any suggestion/advice or doesn’t our customer need to fill in a technical form? Looking forward to your response. Kind Regards
Asked on Nov 03 2011
Thank you very much for contacting us with the concern of this customer. Indeed, the picture on the website is not a WEstern blot, however a SDS page where the 3ug of the protein have been loaded and than stained with coomassie blue. If the customer gets only a faint band with a coomassie blue staining, I think this is expected if only 0.1ug are loaded. It is 30 times less. If the customer however is running a Western blot, where the protein is detected with an antibody, I would expect a stronger band if the antibody is working well. Please let me know if the customer is running a Western blot, in that case I would recommend to fill the questionnaire so I can try to help to troubleshoot. If the customer is concerned that not enough protein is in the vial, I would recommend firstly to measure the concentration of the protein. This can either be done by a BSA assay, OD measurement or running a protein sample of known concentration (e.g. BSA) as a standard on the SDS Page. I hope this information is helpful. Please let me know if you have any further questions.
Answered on Nov 03 2011