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
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. :)
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?
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.
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
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
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?
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.
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.
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
Never ever stop making videos. Thank you so much.
How can you be always so clear? I love your videos!
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!!
Thank you, it was a good explanation. Please keep working, there are so many people that need it.
What a clever way to distinguish between different types of proteins
This is very helpful for helping me understand material in my molecular biotechnology class.
This man deserves my degree.
very helful videos ... thank u ... simple explainations to tough topics....
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
It is just unmatched... Thank you thank you thank you a million
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
Amazing explanation I understand everything with your Videos!!
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
Congrtulations for your great work and easily understandable teaching !!!
Best explanation of 2D-PAGE!!
you are an absolute legend man
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
Very Helpful.. Thank you so much sir..
Where will I be without your videos? Thank you !
I ould have failed without this channel and the channelShomu's Biology
This is so awesomely exlained.. Thank you very much!!!
Wonderful lecture finally understood this
Excellent video
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 very much sir 💝
My concept is clear
Love from INDIA # BHARAT
Very helpful , thanks a lot teacher.
Thank you so much! very clear explanation!
tammy wong you're welcome Tammy :)
Omg! You are a life saver! Thank you so much!
Super explanation sir
You Are the BEST! ALWAYS!
This is brilliant. Thank you.
thank you so much, because this video I really understand this topic.😊
I love the way you discuss.. by saying "We" hehe
you made it too easy.. thanks
Thank u this video very helpful
You are the best , thank u so much , your videos are so so helpful :)
+okba zahra hi:)
+Felipee Rincon hello :)
very helpful, thank you so much!
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?
I learned a lot. Thanks!
It can Not be better, thanks a lot ❤
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
You are AMAZING! THANK YOU!
very clear, Thank you VERY MUCH.
very helpful... Thankyou
Thank you, useful.
excellent
I love this. I love you. Thank youuuu!
Are proteins taken out of the pH gradient gel to apply SDS or is this done inside the gel?
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
Amazing!
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
i still dont get it
Great✨
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?
Respect ✊🏼
you saved me from my supervisor !
Is this Native PAGE electrophoresis?
Thank you !!
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Yo, thamk you very much mister
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.
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.
Create your own religion! This got to be scientifically religious the way you explain so clearly. Thanks
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nice done (Y)
So those people who disliked this video think they can make a better lol
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.
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