Nylon membranes - (Mar/02/2006 )
Hello, I am trying to set up to do Western blotting for my boss and he has a bunch of Boehringer mannheim nylon membranes and some Genescreen and genescreen Plus nylon membranes. Can I use these for protein transfer? He has a bunch from when he did Northerns. The Bio-Rad mannual says PVFD's (Polyvinylidene difluoride) but the paper whose experiment we want to recreate just says that the gels were opposed to nylon membranes. I wasn't sure if the membranes I have were only for DNA and RNA or if they will work with proteins. I would like to save money if I can use them. Any help would be appreciated.
P.S. We always used nitrocellulose when I was in school so I don't know very much about other membranes.
the best way to answer this, would be to go to the manufacturer's site and look up product specs for each type of membrane
ex, genescreen; go to Schlier and Schuell (I'm sure I've spelled it wrong 
and look up specs for that type to see relevant applications for each membrane
I never used nylon membrane (only nitrocellulose and PVDF),
but it seems you can use nylon, I've seen it on this flier :
http://content.labvelocity.com/protocols/5...cycling_Kit.pdf
PS : Schleicher and Schuell
ex, genescreen; go to Schlier and Schuell (I'm sure I've spelled it wrong
and look up specs for that type to see relevant applications for each membraneI have and from what I have found they are most often used for DNA and RNA, but I was just wondering if any one knew why they couldn't be used for Proteins. All the information I douns discussed using them for Northern and Southerns.
they can be used for protein. you have to block exhaustively or you will get a high background.
So then a standard 5% milk blocking solution won't be sufficient, or would I just have to let it block longer?
I don't have a lot of experience with milk, we use bsa and normal goat serum for blocking but we use stronger block and longer time (sometimes overnight at 4C).
Ok Thanks for the info guys, plus I found a 0.45 micron membrane stashed away that specifically says it can be used for proteins and DNA so I will use that. Thanks again for all your help.
Actually I had another membrane question. What is the average shelf life for a nylon membrane? How old can they be before they probably shouldn't be used? The ones i found are from Feb.....of 1990 
I'm not aware of any hard set expiration time for membranes. a lot depends on storage conditions.
you can always try it with a test sample.