Choosing a gel percentage for gel electrophoresis (polyacrylamide & agarose)

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Use a higher percentage gel to separate smaller molecules and lower percentage gel to separate bigger molecules. Have some of both? A gradient gel might serve you well! Not sure what to use? Look to a migration chart to tell!
ThermoFisher Gel Electrophoresis Tables: www.thermofisher.com/us/en/ho...
more on agarose gels: bit.ly/agaroses ; RU-vid: • Agarose science - comp...
more on polyacrylamide gels: bit.ly/polyacrylamidepolymeri... ; RU-vid: • Casting polyacrylamide...
more random practical lab tips: bit.ly/lab_tricks_page

more about all sorts of things: #365DaysOfScience All (with topics listed) 👉 bit.ly/2OllAB0 or search blog: thebumblingbiochemist.com

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27 фев 2023

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Комментарии : 4

@bozfamily10 Год назад

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?

@thebumblingbiochemist Год назад

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!

@bozfamily10 Год назад

@@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!

@thebumblingbiochemist Год назад

Feel free to let me knnow how it goes! Good luck!