Dear Artem

the problem is that the range of the temperature control of the most common plate readers is very restricted (generally do not work at temperature higher than 45-50°C) probably because at this temperature the sample evaporation become an issue.

To my knowledge the only alternative to RT-PCR is single cuvette thermo-regulated fluorimeter but in this case you lost one of the most advantages of DSF that is the throughput.

However if you have just to analyze few samples and you can use a larger sample volumes, it can be done.

Using a single cell fluorimeter with thermostatic regulation i was able to generate the curve reported in Fig5S of Banci L, Bertini I, Ciofi-Baffoni S, et al. Human Sco1 functional studies and pathological implications of the P174L mutant. Proc Natl Acad Sci U S A. 2007;104(1):15-20. doi:10.1073/pnas.0606189103

https://www.pnas.org/content/pnas/suppl/2006/12/14/0606189103.DC1/06189Fig5.pdf

p.s; if you are interested in my blog ProteoCool https://proteocool.blogspot.com/

(Differential Scanning Fluorimetry - DSF:A simple and powerful technology for protein QC)

best regards

Manuele

Dear Manuele,

Thanks for your response and links.

I understand that most plate readers work at temp. range up to 45-50°C. That is why I specifically wrote that plate reader in our facility has a specific factory-installed heating module and the max. working temperature is 65°C.

And the figure from your PNAS paper implies that it is possible to reproduce the desired curves in microplates (because the temp. scale is also up 65°C, and our protein has a melting temperature around 50 deg, so should be enough).

p.s. your blog is really cool and illustrative, thanks a lot

Best,

Artem