immunolocalization q's

Robin L. Wright wrightr at ZOOLOGY.WASHINGTON.EDU
Mon Aug 22 19:36:32 EST 1994


We use 1% polyethylenimine to stick down fixed yeast cells for  
immunofluorescence.  The cells really stay stuck down all the way  
through the entire process.  The only problem is that you may put  
down too many cells if you are used to using polylysine.  The  
concentration of cells on the slide at the beginning should be the  
same as you need to see at the end of the process.  Definitely don't  
put down more cells than you need, just to account for loss of cells.

Protocol:  Filter 1% polyethylenimine through 0.22 micron filter.   
Place drop in well of slide.  Wait 1-2 min.  Rinse off with distilled  
water and air dry.  Add cells and let settle for 5-10 minutes (in a  
moist chamber).  Continue as usual.

Robin Wright



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