Hi! Love your videos; they're helping me in the biology part of my Ph.D.! I am working on siRNA-antibody conjugates, and I am trying to separate the unconjugated siRNA/antibody and conjugated molecule. It looks like a gradient gel would be best, but I am still worried about the separation between the unconjugated antibody and the conjugated antibody. Any tips in that regard?
Thanks so much! I'm not sure how well a gel would be able to resolve that. Have you tried running a gel? Do you see multiple bands? Rather than a gradient, you'd probably have better luck if you tailored it around the size of the antibody for best resolution there. If you don't have success with the gel, you could maybe try doing an affinity capture with biotinylated DNA complementary to your siRNA sequence? Best of luck!
@@thebumblingbiochemist Thank you for the response! I am in the process of getting materials to run a gel. A previous student used ultrafiltration to separate larger proteins from siRNA and did two separate gels, but I am not convinced it will work in my case. I will check out affinity captures! Thanks again!
Hi! Love your videos; they're helping me in the biology part of my Ph.D.! I am working on siRNA-antibody conjugates, and I am trying to separate the unconjugated siRNA/antibody and conjugated molecule. It looks like a gradient gel would be best, but I am still worried about the separation between the unconjugated antibody and the conjugated antibody. Any tips in that regard?
Thanks so much! I'm not sure how well a gel would be able to resolve that. Have you tried running a gel? Do you see multiple bands? Rather than a gradient, you'd probably have better luck if you tailored it around the size of the antibody for best resolution there. If you don't have success with the gel, you could maybe try doing an affinity capture with biotinylated DNA complementary to your siRNA sequence? Best of luck!
@@thebumblingbiochemist Thank you for the response! I am in the process of getting materials to run a gel. A previous student used ultrafiltration to separate larger proteins from siRNA and did two separate gels, but I am not convinced it will work in my case. I will check out affinity captures! Thanks again!
Feel free to let me knnow how it goes! Good luck!