Omg your lectures are awesome! Ive been watching all the gel electrophoresis videos and its so clear and also in details i dnt have to waste all my time looking for info on the internet anymore. Thank you!!
simply thank you, you are the best teacher in the universe, you make things so easy , your every vedio is amazing!!, thank you for making us so knowledgeable
It's a technique of separation and visualization, but not used as a method of purification, as purification implies that the protein is in it's native conformation. SDS denatures all of the proteins present so that they could not retain any activity and lose their binding properties.
Im assuming after you run the 2D gel, you can then do a westernblot as you would with a typical SDS gel and then treat with antibodies to determine different post translational modifications of a protein for example?
Question (might be stupid but ok xD) : with the 2D gel electrophorisis can you find out how many polypeptides,from each protein, there are at your first solution? No right? You just separate them based on these two different properties
Thank you for this. You do a much better job at explaining this than my professor. Likewise with a few other videos you have made. I just subscribed and gave your videos a thumbs up. :)
wouldn't the SDS apply a strong negative charge on all the proteins and affect horizontal motion towards the negative end in the Isoelectric focusing component? Assuming both SDS-Page and Isoelectric focusing is occurring together.
Thank you very much for your video. I have a quick question, when the pI is added to the SDS page, will the mass and size increase as you’re going down the SDS page?
i have a presentation on isoelectric focusing on tuesday. i face problems in some points i cannot understand immobilized ampholytes, 2D electrophoresis is type of isoelectric focusing capillary method and SDS are also part of it. how to load sample in gel can you explain me
But how do we separate those proteins which may have the possibility of having the same PI value as well as the same molecular mass. I.e they can't be distinguished by 2d technique?? Please advise.
You can´t IP separation must be done first, because it separes based on the IP so you want the protein to be on its natural form and with it´s natural charge (+ or-), once you do SDS you denaturate proteins wich makes them to lose their dimensional conformation, they are long chains and SDS wich is negative binds to them, giving all proteins a negative charge and thats why now you can separate them ut of their size instead due att all of them have the same charge, wich is negative. So IP first then SDS always.
I wonder how you denature proteins after isoelectric focusing to work in SDS-page, if you take gel from first step the proteins are in their isoelectric point and wont move.
One of the best explanations Ive heard on youtube for any molecular biology technique. Great Job!
Love you. Just pure brilliance across such a vast field. Bravo!
the main objective of all the aklectures videos is to get all your basics and concepts clear
How can you be always so clear? I love your videos!
Never ever stop making videos. Thank you so much.
This man deserves my degree.
you are an absolute legend man
Omg your lectures are awesome! Ive been watching all the gel electrophoresis videos and its so clear and also in details i dnt have to waste all my time looking for info on the internet anymore. Thank you!!
What a clever way to distinguish between different types of proteins
Best explanation of 2D-PAGE!!
This is very helpful for helping me understand material in my molecular biotechnology class.
A youtube science teacher without an asian accent with a clear and concise explanation?
I must be dreaming..
He realised there was a market and he is taking full advantage of it :D
Thank you, it was a good explanation. Please keep working, there are so many people that need it.
simply thank you, you are the best teacher in the universe, you make things so easy , your every vedio is amazing!!, thank you for making us so knowledgeable
It is just unmatched... Thank you thank you thank you a million
Amazing explanation I understand everything with your Videos!!
It's a technique of separation and visualization, but not used as a method of purification, as purification implies that the protein is in it's native conformation. SDS denatures all of the proteins present so that they could not retain any activity and lose their binding properties.
U have no idea how much You´ve been helping me !! Thank You soooo much ! Since I found you, I fell in love with Biochem again lol
Thank you very much sir 💝
My concept is clear
Love from INDIA # BHARAT
Congrtulations for your great work and easily understandable teaching !!!
very helful videos ... thank u ... simple explainations to tough topics....
Where will I be without your videos? Thank you !
I ould have failed without this channel and the channelShomu's Biology
Thank you for this clear teaching. This video helped me for preparing my exam :)
Which exam ?
@@Raka96876 Genomics and proteomics
@@yaseminacar4329 Are you doing Ph.D in genomics ?
@@Raka96876 it was my undergraduate exam. I am doing PhD in molecular pharmacology
Wonderful lecture finally understood this
Excellent video
Very Helpful.. Thank you so much sir..
This is so awesomely exlained.. Thank you very much!!!
You Are the BEST! ALWAYS!
Omg! You are a life saver! Thank you so much!
thank you so much, because this video I really understand this topic.😊
Thank you so much! very clear explanation!
tammy wong you're welcome Tammy :)
Super explanation sir
Very helpful , thanks a lot teacher.
This is brilliant. Thank you.
I love the way you discuss.. by saying "We" hehe
Im assuming after you run the 2D gel, you can then do a westernblot as you would with a typical SDS gel and then treat with antibodies to determine different post translational modifications of a protein for example?
Question (might be stupid but ok xD) : with the 2D gel electrophorisis can you find out how many polypeptides,from each protein, there are at your first solution? No right? You just separate them based on these two different properties
Thank u this video very helpful
Create your own religion! This got to be scientifically religious the way you explain so clearly. Thanks
you made it too easy.. thanks
It can Not be better, thanks a lot ❤
Thank you for this. You do a much better job at explaining this than my professor. Likewise with a few other videos you have made. I just subscribed and gave your videos a thumbs up. :)
Thank you, useful.
excellent
very clear, Thank you VERY MUCH.
You are the best , thank u so much , your videos are so so helpful :)
+okba zahra hi:)
+Felipee Rincon hello :)
You are AMAZING! THANK YOU!
wouldn't the SDS apply a strong negative charge on all the proteins and affect horizontal motion towards the negative end in the Isoelectric focusing component? Assuming both SDS-Page and Isoelectric focusing is occurring together.
They r done seperately
Nope... they aren't been done together...first you do the isoelectric focusing and then SDS page
I learned a lot. Thanks!
very helpful, thank you so much!
Are proteins taken out of the pH gradient gel to apply SDS or is this done inside the gel?
Respect ✊🏼
Thank you very much for your video. I have a quick question, when the pI is added to the SDS page, will the mass and size increase as you’re going down the SDS page?
very helpful... Thankyou
I love this. I love you. Thank youuuu!
you saved me from my supervisor !
i have a presentation on isoelectric focusing on tuesday. i face problems in some points i cannot understand immobilized ampholytes, 2D electrophoresis is type of isoelectric focusing capillary method and SDS are also part of it. how to load sample in gel can you explain me
Great✨
Amazing!
Is this Native PAGE electrophoresis?
Thank you !!
But how do we separate those proteins which may have the possibility of having the same PI value as well as the same molecular mass. I.e they can't be distinguished by 2d technique??
Please advise.
This is to check stream purity. For separation, I would recommend using them in chromatography and looking at affinity chromatography in particular.
can you reverse the steps and perform mass separation first and PI separation second. Why or why not?
You can´t IP separation must be done first, because it separes based on the IP so you want the protein to be on its natural form and with it´s natural charge (+ or-), once you do SDS you denaturate proteins wich makes them to lose their dimensional conformation, they are long chains and SDS wich is negative binds to them, giving all proteins a negative charge and thats why now you can separate them ut of their size instead due att all of them have the same charge, wich is negative. So IP first then SDS always.
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i still dont get it
I wonder how you denature proteins after isoelectric focusing to work in SDS-page, if you take gel from first step the proteins are in their isoelectric point and wont move.
So those people who disliked this video think they can make a better lol
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