If you are analyzing the thermal shift of your protein samples using differential scanning fluorimetry (DSF), then you can use the MS Excel based tool which is described here http://thermofluor.org/resources/DSF-Analysis-Manual-v3.0.pdf and the files can be downloaded from ftp://ftp.sgc.ox.ac.uk/pub/biophysics

Thank you, I'll try it.

HI,

Both of these links now lead to dead ends, does anyone know where I can get now get hold of the spreadsheet and manual?

Many thanks,

You can access the previous site going to http://www.beta-sheet.org/

Thanks Boopathi and Ricardo

Hi, since it is a ftp site, ResearchGate may fail to redirect when you click the above link. Instead, copy the link, open a new browser window, paste the link and hit enter. It should work. Cheers!

 Cannot access ftp://ftp.sgc.ox.ac.uk/pub/biophysics

Can some one send me the file on my email: [email protected]

Thanks 

Deepak Gurbani

I would suggest you to use protein thermal shift software 1.3....

A number of programs out there - here are some that I know about:

Meltdown (Article Meltdown

DMAN (https://academic.oup.com/bioinformatics/article/28/3/439/188888)

MTSA (Article MTSA—A Matlab program to fit thermal shift data

SimpleDSFviewer

The 5-parameter sigmoid fitting from the MTSA program is also available as an online JavaScript application: http://paulsbond.co.uk/jtsa

You are saying you don't even know if these assays will generate good results and you have no experience in this technique. Therefore, the first thing to do is running a few tests first. Just a visual inspection will tell you if they are similar to other reported similar assays.

After that, I do not think you need specialized software. You need the raw data for the fluorescence intensity (FI) as a function of temperature (T), which should be readily obtained from the instrument.

Then, you can plot FI as a function of T. If you get the usual unfolding trace, the Tm can be estimated from the point of maximum slope or inflection point. In fact, the Tm does not correspond to the temperature for maximum slope (Tmax,slope > Tm, if the heat capacity of unfolding is not zero), but it is reasonably close. Therefore, in order to calculate the position of the inflection point you can calculate the first derivative of the unfolding trace (dFI/dT vs. T) and locate the maximum, or calculate the second derivative (d2FI/dT2 vs. T) of the unfolding trace and locate the intercept with the T-axis.

Of course, doing this for s single curve is easy, but for a set of several curves may be cumbersome. However, it can be automated in any data analysis software package.