At the protocol I'm using now there is an added note indicating that it is possible to freeze the protein samples mixed with the laemmli to temporarily store them at -20ºC and denature them before performing the western blot. However, one of my coworkers says she tried it once and it didn't work out. ¿Can anyone please tell me if they have done this before and how it turned out? I have a large number of samples and this could save me some time. Tips and suggestions are appreciated.
I would suggest take a few microlitres of protein sample and mix with same amount of buffer and proceed to store at -20 to be denatured later.
In case someone is interested, I tried and it worked quite alright. I froze the amount of protein I needed with the corresponding laemmli at -20ºC and denatured it before the WB. Also, I did it using both laemmli prepared at the laboratory (with beta mercaptoethanol) and commercial laemmli, and I found no difference.
- Badges
- Science method