Well it's finally done. After over 4 months of failed cloning attempts and protease (proteins that chew up other proteins) problems, my protein is finally purified and ready to be injected into 2 bunnies who will hopefull have a huge immune response to my protein and give me lots and lots of antibodies against it! With luck, I will have my antibody in just 4-6 weeks!!
I have learned quite a bit of biochemistry during this whole protein purification process. At least the failures have helped me to gain experience in troubleshooting and thus in number of additional biochemical assays. Now my resume/CV can include doing the following:
Making/Running SDS-PAGE gels
Western blots
GST-protein purification
His-tag protein purification
Performing Bradford tests
Protein induction
Making Competent cells
I guess I learned a lot this semester. I also met my own personal deadline-to have the protein purified by the end of the semester. I'm 2 days ahead of schedule.
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