Storing tissue for protein and RNA extraction? - (Feb/19/2008 )
I have a bit of problem getting good results for my Western blot...i.e. sometimes I get a good band for the protein of interest, other times, following the same steps etc, I get poor signal. I store my tissue sample post snap freezing at -80 Degree C and was wondering if that is sufficient. Another person in the department said he stores his samples in Liquid nitrogen for tissue intended for RNA and protein extraction.
I was wondering if anyone can comment on what storage condition to use to achieve good quality RNA and protein samples. We do not have liquid nitrogen facility in my lab.
I store plant material at -80ºc and it's fine for a long time (we don't have liquid nitrogen storing facilities too).
Thanks... that is good to hear! Just wondering if there is a different in storing mamalian versus plant tissues...
Can't help you there - I only work with plants...
Nope, no for me though. I store my protein extracts at -20 and they are still good after.. let's say several months. Maybe you problem resides more in the freeze/thaw than in the protease contamination..
I don't freeze thaw samples... they are stored in small aliquotes and once thawed, they are used and remaining samples discarded...
Any other comments/suggestions...
Hi skll, even i face the same problem. Even though i dont freeze thaw the sample and store at-80 c, it gets denatured.
-80'C. best option.
Thanks V and vinuuuu..
It seemed that most people would store their tissue and protein samples at -80DegC. Anyway, what about RNA samples?
vinuuuu: how do you overcome your protein degradation problem... I suppose if you store at -20DegC... it is a non- frost free freezer... as frost-free freezers tend to cycle their temperature, sometimes to about -10DegC... and hence can cause freeze thawing effect at times...
RNA is stored at -80'C.
protein degredation can be stopped by using protease inhibitors... usually in a cocktail. tablets can be disolved in water, and then added to the lysis buffer you're using.