Long-term storage of Westerns? Freezing?

Jayakumar, R R.Jayakumar at RoswellPark.org
Thu Dec 2 11:26:32 EST 2004


Dry them before freezing in -80C.  PVDF membranes can be dried by floating them on methanol for 15 seconds (they will sink into the methanol eventually) and leaving them at room temperature for 15 minutes.  The dried membranes shoudl be wrapped in tissue wipes and again sandwiched between two stiff filter paper folder (use extra thick whatman filter paper)before placing them inside a flat cryocontainer. They become extremely brittle when frozen.  Thaw them at room temperature before even attempting to remove them from the filter paper. Dried membranes cannot be stripped and reprobed, since drying causes the bound antibodies (which are proteins themselves) to bind very strongly to the membrane.  If you need to reprobe them, then strip them first before drying them.  You can do the blocking immediately before resuse.
   
Jayakumar

-----Original Message-----
From: owner-methods at hgmp.mrc.ac.uk
[mailto:owner-methods at hgmp.mrc.ac.uk]On Behalf Of Wolfgang Schechinger
Sent: Thursday, December 02, 2004 7:44 AM
To: methods at hgmp.mrc.ac.uk
Subject: Re: Long-term storage of Westerns? Freezing?


It seems to possible to store them dried between 2 sheets of paper or in a
small sealable plastic bag. I'd recomment a desiccator. If you freeze them,
condensing water and ice might cause problems.

Wo

At 14:26 22.11.2004 -0800, you wrote:
>Hello,
>Can Westerns be stored frozen?
>Before using?
>After probing? And if so, is stripping recommended prior?
>
>Please send response to candyman476 at netzero.net
>
>THanks!
>
>
Wolfgang Schechinger, PhD (Dr. rer. nat.)
Chemistry and Life Sciences

BGFA XU44
Med Clinic I / Endocrine Research 
University Hospital "Bergmannsheil"
Bürkle-de-la-Camp-Platz 1
44789 Bochum
Germany

Phone +49-234-302-4789
Fax   +49-234-307-4790

wolfgang.schechinger at gmx.net




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