🧪Thermal Denaturation of Proteins using Circular Dichroism || Practical Biochemistry 

It's for the main lamp, otherwise it will get damaged

What changes in CD spectrum do you see?

Hello. What kind of aggregates are they? Amorphous or like fibrils, and how would you describe the data you are getting? Are you getting a lot of noise on the left side of the curve?

@@biochemistmelo Thank you so much for responding! This is awesome! They are amorphous and the graph starts at zero (right side) and then keeps going down (-1 mdeg) from 230-215nm (-2 mdeg) at 210 and goes up (-1mdeg) and the 205 nm goes down (-4 mdeg). Yes! It looks like a lot of noise on the left side of the curve as you mention!

@@nicholemontero8481 can I also ask what you dialyzed the samples in? And concentration of sample

@@biochemistmelo Of course, I used 20 mM NaH2PO4, 0.5 M NaCl! sample was a little low 3.56 µM

@@nicholemontero8481 yes both the NaCl concentration is really high and the protein is low (try 5x more). I would reduce the salt as much as possible if it doesn't affect your protein.

As a graduate student it's been a long time since I had an actual text book to read. I usually read many journal papers and manuals to construct an understanding of a technique.

So there is a baseline for when the protein is still folded and when its completely denatured with a slope in between. The midpoint of the slope is Tm. For example if it starts unfolding at 30 and finishes at 50, the Tm is 40. The cooperativity comes from how steep the slope is. If you have something that's almost a sharp 90 degrees and another that's 30 deg, then the sharp transition is the more cooperative unfolding. If you have more than 2 datasets then measuring the slope (difference in y/x). I suggest starting at a really low temp like 10 and ending at a high temp like 80 so you can clearly see the baseline before and after the main slope of transition.

Yes, what are your interests?

I m studing general up to now And want to works on health biotech I need guidance what ,scholarships and fellowship for future studies

@@aimakhan976 the scholarships really depend on what country you want to study in. I studied in Canada.

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