Maa'm after the 1D sepration of proteins ..they were at their isoelectric point ...so had zero chagre But in second sepration i.e. SDSsepration u said proteins move from negative to positive electrode If they didnt had any charge on them ....how are proteins moving For sds we need charge on proteins to move in electrical feild Please explain this
Thanks for the video; what is the Conc. of the SDS PAGE gel, and is it only resolving without stacking? is there any buffer added with sample during separation of the PI step?
After performing IEF the proteins will become neutral, will there be movement after applying SDS which obtains -ve charge due to the presence of ampholytes.
Hello, no actually you cannot perform SDS before you should keep the same order .. The idea is to separate proteins who have the same MW ... if you perform SDS first then you have your proteins in the gel ... so you need a special container to put the gel in and to apply the PH gradient ... but in this case, you perform the IEF when the proteins are in the liquid and then you apply the strip directly to the gel ...
Do you have any lecture on Mass spectroscopy??please let me know!! And if not then a humble request to you make lecture on that topic please !!! And its a best lecture on this particular topic which is really helpful to understand !!!Thank you so much Mam!!👏👏👏🙏
Why haven't this video more views or comments?! I already sad "thank you" in your video about SDS-Page, but I'll say again: THANK YOU SO MUCH! Please don't stop making videos
Hello, actually I am working at the moment on a huge topic which is the CRISPR-Cas9 technique, which needed tow videos to be prepared, I will upload the videos soon and then the next will be blotting technique, and then I will work on chromatography because it might also need tow videos. stay around my friend, a lot of interesting videos are coming up ;)
Hey ... Here are the two videos about blotting techniques, the first is about western blotting and the second is about southern blotting and northern blotting: ua-cam.com/video/BguNIPiCmv8/v-deo.html ua-cam.com/users/edit?o=U&video_id=RvbzjYM_Ok0 I hope you will enjoy them :)
In the first step, and to separate the proteins according to their PH we dont apply then in a well, we apply the proteins mixture in the container where there is a strip containing a PH gradient, and then the proteins will be separated according to their isoelectric point (see from 7.34 min)
However, I guess the example of the second dimension of separating proteins of different PIs yet with the same molecular weight had a mistake in the figures:)
what i couldnt understand in a full week i got it in 10 minutes, god bless you
Voice is calming, The teaching is clear. thanks alot
i ve just discovered this channel and its amazing
This video was very helpful. Please continue posting such videos. You are a great help
Please make more videos. I love your explanation. It's very thorough 👍🏼
Thank u ma'am for such a wonderful video.very educational and clear
Thank you for your comment .. stay tuned ;)
Maa'm after the 1D sepration of proteins ..they were at their isoelectric point ...so had zero chagre
But in second sepration i.e. SDSsepration u said proteins move from negative to positive electrode
If they didnt had any charge on them ....how are proteins moving
For sds we need charge on proteins to move in electrical feild
Please explain this
Thanks for the video; what is the Conc. of the SDS PAGE gel, and is it only resolving without stacking?
is there any buffer added with sample during separation of the PI step?
Mam what if the two proteins have same isoelectric point and molecular weight??
wonderful. without this video I could not understand it. Thank you very much
This video is clear and very very useful. Thank you!
Very clear video for beginners
at ph =3 what will be the emergences order of the amino acid in column on the basis of defferent pka value ?
I am taking a course in biochemistry titled biochemical reasoning. Do you have any material on it?
Thank you, your video are extremely helpful. Please don't stop making them.
After performing IEF the proteins will become neutral, will there be movement after applying SDS which obtains -ve charge due to the presence of ampholytes.
Extraordinaryly explained madam thank u so much
where can I find articls which use this method!?
Excellent video! Congratulations
Excellent explanation. Thanks
I loved this video. Thank you so much !! Eres la mejor :D
Very nicely explained ❤️
Hello, I wanted to ask can the SDS PAGE be performed prior to the IEF? Why do we need to maintain the order of IEF and then SDS-PAGE? Thank you!
Hello, no actually you cannot perform SDS before you should keep the same order .. The idea is to separate proteins who have the same MW ... if you perform SDS first then you have your proteins in the gel ... so you need a special container to put the gel in and to apply the PH gradient ... but in this case, you perform the IEF when the proteins are in the liquid and then you apply the strip directly to the gel ...
your accent is very clear and really thanks for your explanation
Do you have any lecture on Mass spectroscopy??please let me know!! And if not then a humble request to you make lecture on that topic please !!! And its a best lecture on this particular topic which is really helpful to understand !!!Thank you so much Mam!!👏👏👏🙏
Why haven't this video more views or comments?! I already sad "thank you" in your video about SDS-Page, but I'll say again: THANK YOU SO MUCH! Please don't stop making videos
Thank youu for the encouragement, I will not stop :)
Hi
thank you ! really helpful during this pandemic situation.
Really really thank you so much! Your videos makes everything so simple, they're so useful :) ❤️
It was very helpful. Thank you very much!!!
Madam when is next video coming? I asked for Blotting techniques and Chromatographic techniques.
Hello, actually I am working at the moment on a huge topic which is the CRISPR-Cas9 technique, which needed tow videos to be prepared, I will upload the videos soon and then the next will be blotting technique, and then I will work on chromatography because it might also need tow videos.
stay around my friend, a lot of interesting videos are coming up ;)
Thanks, I'm eagerly waiting for the videos to come. :)
Hey ... Here are the two videos about blotting techniques, the first is about western blotting and the second is about southern blotting and northern blotting:
ua-cam.com/video/BguNIPiCmv8/v-deo.html
ua-cam.com/users/edit?o=U&video_id=RvbzjYM_Ok0
I hope you will enjoy them :)
Really ... can u please tell me about the mistake!!! In which video is it ... the agarose gel electrophoresis???
this should have more views. thank u for putting this up it was really helpful!!!!! :-)
plz mam can you post about pulsed field gel electrophoresis and native too coz i hav got exams soon bt i didnt get it😐😐
I will try to make it .. but tell me .. when is your exam ??
Hei, a bit late but here you are, the link for the pulsed field electrophoresis video:
ua-cam.com/video/__QhCX12h8I/v-deo.html
Thanku mam for this nice vedio.Mam please says about denatured and non denatured 2D gel.
when we load the protein in the well ,then how it is seprate to its ph
In the first step, and to separate the proteins according to their PH we dont apply then in a well, we apply the proteins mixture in the container where there is a strip containing a PH gradient, and then the proteins will be separated according to their isoelectric point (see from 7.34 min)
So how do we denature the proteins before loading it onto to the SDS page gel? Do we just flood the ph strip with loading dye?
thank you so much it's very useful
well done! it is very useful
Thanks a lot madam
Soo good ...but mam plzz.speak english like india for easy to listening for indian
we know the ph of protein before loading
Normally, we know the isoelectric points of the protein of interest ...
Thanks doctor this is amazing
However, I guess the example of the second dimension of separating proteins of different PIs yet with the same molecular weight had a mistake in the figures:)
you are the best
Mam please upload for immuno gel electrophoresis....i understood only because of this. Thank you so much
Great explanation! thank you
Thank you
If you please also upload the ppt files that will be great
Thank you so much
Ver, very useful!!!!
Thank you!
Thank you❤️
v good..
you are so clear!
thank you!
👍👍👍
THANK U
thanks for the explanation. but 15 min is too long
Thanks doc 💗
Thank you