You don't understand how valuable of a resource you are! You break things down in such a way the viewer can easily grasp but, you know it took you countless restless hours to decipher all this information. There's not many people like you but, only a few chosen people have the opportunity to discover you. Keep the content flowing; your channel is a hidden gem I hope no one else stumbles upon because I want to hoard all your content to myself. lmao
Hii i have a few questions 1)can i use dailysis bags instead of dailysis cassettes ? 2) can dailysis bags of 14000D be used to do dailysis of protein with 350000 D? 3) And if all the above questions answer yes then what should be the exact time for me to keep the solution for dailysis? Like for how much time the dailysis should be done...?
Hello! Thanks for the video, it was very clear. I have a question, where can i find the protocol to concentrate proteins with Polyethylene glycol? Did you do it? Or you read about it in a book? That information would be very useful to me :)
@@thebumblingbiochemist I've already tried :(, but I didn't find anything about concentration of proteins. The results are about the polyethylene glycol as a drug or to precipitate proteins with it. I don't know which concentration of PEG use or how to make the solution: in water, in the same buffer of my protein? I have many doubts and I didn't find this method in the books that I have. I know there are so many ways to concentrate the proteins but we don't have enough money to buy the reagents. However, the laboratoy next to us have PEG and when you mencionated it y thought to use it. So, if you have any other information I would be so grateful. I am from Argentina by the way. Thank you
Many thanks. How to do the operations from minute 17 in a sterile way : soaking of membrane, etc and the liquid from 2 liters Berzelius to remain sterile. I need an idea . Good work. Many thanks for demonstration.
At the final of dialysis , how to concentrate ( a simple Method) the proteins ? Is It possible by a simple centrifugation at maximum 10,000 g followed by a supernatant removal ? I mention the dimension of protein is under 0.2 microns. Thanks
You don't understand how valuable of a resource you are! You break things down in such a way the viewer can easily grasp but, you know it took you countless restless hours to decipher all this information. There's not many people like you but, only a few chosen people have the opportunity to discover you. Keep the content flowing; your channel is a hidden gem I hope no one else stumbles upon because I want to hoard all your content to myself. lmao
Thank you so much! But the great thing about this is it's free for anyone and people watching doesn't steal it from others :)
You are like the lab senior that I never had in my graduate school! Many thanks for the live demonstration
Happy to help!
Just wanted to tell you that this video rescued me while I had to purify a protein urgently but I had no experience with the dialysis. Thank you!
So happy I could help! And hope the purification went well
Thank you so much, it was a lot easier to understand the process (and also the practical aspects of it) after watching this demonstration.
Glad it helped!
Blessed we are to have such useful information.
I tried the dialysis casette but for some reason I could not retain my protein. What are some common mistakes that can lead to this?
Do you mean it's leaking out? Or dialyzing through? Or crashing out?
Thank you for the resourceful information. I really appreciate
You are quite welcome!
Thank you for all these videos
Glad you find them helpful!
good stuff! I like these detailed videos :)
Thank you!
Hii i have a few questions 1)can i use dailysis bags instead of dailysis cassettes ?
2) can dailysis bags of 14000D be used to do dailysis of protein with 350000 D?
3) And if all the above questions answer yes then what should be the exact time for me to keep the solution for dailysis? Like for how much time the dailysis should be done...?
Yes and yes. And typically a couple of hours, then change out then buffer, then overnight
@@thebumblingbiochemist one last question is it necessary to buffer like only dailysis with water won't be enough?
Correct. You want the dialysis fluid to be the buffer you want your sample to be in.
thank you, I was struggling to understand it
Happy I could help!
Hello! Thanks for the video, it was very clear. I have a question, where can i find the protocol to concentrate proteins with Polyethylene glycol? Did you do it? Or you read about it in a book? That information would be very useful to me :)
Thanks! I haven't personally done it sorry, but am sure if you Google you can find one. I
think you just add PEG outside the bag. Good luck!
@@thebumblingbiochemist I've already tried :(, but I didn't find anything about concentration of proteins. The results are about the polyethylene glycol as a drug or to precipitate proteins with it. I don't know which concentration of PEG use or how to make the solution: in water, in the same buffer of my protein? I have many doubts and I didn't find this method in the books that I have. I know there are so many ways to concentrate the proteins but we don't have enough money to buy the reagents. However, the laboratoy next to us have PEG and when you mencionated it y thought to use it. So, if you have any other information I would be so grateful. I am from Argentina by the way. Thank you
Sorry! It should be the same as the buffer you want, but just with PEG added
Many thanks. How to do the operations from minute 17 in a sterile way : soaking of membrane, etc and the liquid from 2 liters Berzelius to remain sterile. I need an idea . Good work. Many thanks for demonstration.
Thanks! I'm not sure the best way to do that other than to make sure to sterilize your work space and all your equipment. Good luck!
@@thebumblingbiochemist Many thanks.
Would using a pump to continuously perfuse fresh solvent reduce the volume of waste solvent? I made 6L of chemical waste before.
I'm not sure how that would reduce waste?
Thankyou so much for make this videoooo 🤗🤗
What about desalting by simply doing spin concentration?
You can use a spin concentrator to desalt, but you'll have to resuspend and re-spin several times.
many thanks for the video!)
Do we need to treat these membranes first with bicarbonate and EDTA ?
I never do
Why to do that ? Which would be the benefits ?
What the red box contains (except the membrane) ?
what red box?
@@thebumblingbiochemist At the minutes 13: 30 appear those red box for dialysis. But what is inside de box ?
just the membrane
At the final of dialysis , how to concentrate ( a simple Method) the proteins ? Is It possible by a simple centrifugation at maximum 10,000 g followed by a supernatant removal ? I mention the dimension of protein is under 0.2 microns. Thanks
No - I suggest centrifugal ultrafiltration (spin concentrators) bit.ly/spinconcentrators & th-cam.com/video/Q_Ua7tIjzWQ/w-d-xo.html. Good luck!