What do you do with YOUR membranes? - (May/12/2006 )
Ok so I have a general question about western blotting. How long is it okay to leave a membrane in TBSt/PBS between stripping/reprobing. For example, after you develop your first blot what do you all do with your membranes. I generally leave them in the saran wrap and put them in the fridge until I need to strip and reprobe. The only thing is, then it runs the risk of drying out which gives you background problems. That said, is it okay to leave it in TBSt in between reprobes? Or is that a problem? I'm a fledgling...help me out!
We throw the membrane after imaging.
i usually store my membranes in PBS at 4C until ready for stripping/reprobing.
i store them RT and do the 4-5 blots as quick as possible. Finally, i wrap them in saran and keep at 4° for future reprobing.
Hi. I usually dry and store my NC membrane in between dry whatman paper at room temperature until they are ready for processing. In between probe, I washed them in TBST and then store them wrapped in saran wrap in the freezer (-20DegC). I dont have any problem with background..have tried this even up to a month. I have also stored them a 4DegC in TBST overnight if I want to reprobe the next day.
Great that helps a lot guys!
...now im trying to redo some of my freind's experiments and thank God she kept the membranes!!!!!
In your experience, how many strips and reprobes can you do before the membrane loses its integrity? I usually hit a wall after 5.