Thanks for the video! Great job, I would just like to add that the value of the titration is that it allows you to determine the binding affinity. You can use the EMSA to determine the Kd (concentration of protein at which half the probe is bound).
Great video! I love your explanations. My professor is referring to your videos for our practicals and they help to understand the principle. Greetings from Maastricht University!
Super nice video! THANK YOU!! I would like to ask you why we need to label our DNA? It is because the DNA itself would be not visible on a NATIVE-PAGE gel autoradiography in the end? And one more question please: In case that I want to check interaction of transcirption factor and promoter region (potential transcription factor binding site) can I also use synthetic DNA seq or i need to extract the DNA and cut it (e.g. with enzymes) and then use it as template for the reaction? I would be very happy to hear your feedback! :)
Thank you ! And yes you are correct. DNA by itself cannot be easily visualized. If you use autoradiography then you would need some radioactive isotope on the DNA for visualization.
Hi, thats an amazing video.. Thanks for all the hardwork that you have put onto it. I am currently conducting an EMSA assay where I observe free DNA decreasing with gradual increase in protein but the Protein-DNA complex does not enter the gel. Also my protein is >450kD so it huge amd multimeric. I am really stuck in here.. Can you give me any ideas that would help me?
@@scigine2378 I use 5% TBE gel.. i have also tried 4%.. I have used 1X and 0.5X TBE too.. I have tried changing the pH of the buffer to 9.. I run the gel at 4°C. I am at a complete loss at what to do..
If you are able to maintain the 4C temperature you can also try increasing voltage. Typically that will make the gel too hot but running at 4C should do the trick
Great watching from Kashmir University
Thank you for the kind words !
Thanks for the video! Great job, I would just like to add that the value of the titration is that it allows you to determine the binding affinity. You can use the EMSA to determine the Kd (concentration of protein at which half the probe is bound).
Great video! I love your explanations. My professor is referring to your videos for our practicals and they help to understand the principle. Greetings from Maastricht University!
Thank you so much for all the work you put into this video! It really helped me. I hope you keep making more videos in the future.
amazing video. you're so clear and thorough in your explanations. thank you!
Thank you for the kind words!
Thankyou :) the video is very helpful 🙏
Pretty nice explanation! Thanks a lot!
Really useful !! Thanks for sharing ❤️❤️
Thanks for the subscribe !
Ur Going good job keep uploading more
Great vid
thank you very useful
I'm glad you liked it! Let me know if you have any questions.
Super nice video! THANK YOU!!
I would like to ask you why we need to label our DNA? It is because the DNA itself would be not visible on a NATIVE-PAGE gel autoradiography in the end?
And one more question please: In case that I want to check interaction of transcirption factor and promoter region (potential transcription factor binding site) can I also use synthetic DNA seq or i need to extract the DNA and cut it (e.g. with enzymes) and then use it as template for the reaction? I would be very happy to hear your feedback! :)
Thank you ! And yes you are correct. DNA by itself cannot be easily visualized. If you use autoradiography then you would need some radioactive isotope on the DNA for visualization.
how titration is done? by the way very through video and in very details.. Thank you
Take a look at figure 1 in this article. A titration is just a concentration dilution. www.ncbi.nlm.nih.gov/pmc/articles/PMC2757439/
Hi, thats an amazing video.. Thanks for all the hardwork that you have put onto it. I am currently conducting an EMSA assay where I observe free DNA decreasing with gradual increase in protein but the Protein-DNA complex does not enter the gel. Also my protein is >450kD so it huge amd multimeric. I am really stuck in here.. Can you give me any ideas that would help me?
Hi! I'm glad you liked the video!
Have you tried reducing the % of the gel? That should increase pore size.
@@scigine2378 I use 5% TBE gel.. i have also tried 4%.. I have used 1X and 0.5X TBE too.. I have tried changing the pH of the buffer to 9.. I run the gel at 4°C.
I am at a complete loss at what to do..
If you are able to maintain the 4C temperature you can also try increasing voltage. Typically that will make the gel too hot but running at 4C should do the trick
@@scigine2378 okay I will try doing that.. And comment here about the results 🐥
Thank you btw for trying to help me out.. You are too kind to be true
Please summarize, you don't need to advertize your other videos in every minute.