Hi all,
I am extracting proteins from cells frozen at -80C. Will it effect the quantity of proteins as compared to the quantity extracted from fresh cell cultures?
Thankyou
I agree with Beverley. Fresh is always best but making lysates from frozen pellets is perfectly ok
Incidentally total amount of protein is not affected by lysing from frozen. Lysing from frozen can lead to protein degradation bit this tends to happen more with older lysates that have been freeze thawed more than 3 x
Freeze thaw multiple times can be used to lyse the cells, but they may not be in buff with proteases inhibitors, but they are frozen so no biological activity or very very little.
There will be some enzyme degradation to protein folding during temperature fluxes and time in cryo.
Total protein will be the same.
Like if you had frozen an egg, not cooked, just frozen. Its all there just not as fresh.. who wants french toast?
Best
The answer is yes, freezing can affect protein quantity, when compared to those in cultured cells, but there are many factors involved, and the degree of effect varies from protein sample to protein sample. Several proteins in cultured cells are in a dynamic state, while those in frozen cells are generally in a state of frozen animation. Therefore, protein quantity is relative to the point of quantitation, not for comparison between frozen and cultured cells. You will need to report the status of cells from which you isolated proteins, to report comparative data.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line