immunolocalization

[mmelan at pattie.wellesley.edu]

Dear Janet,
        I can help you with problem #1 of your inquiry.....
A) You need to be sure that you are using a high molecular weight poly-lysine
        (i.e. greater than 70kD.  We routinely use 150-300kD poly-lysine
        Sigma P1399)  If you use lower molecular weight poly-lysine the sections        will float off of the slides.  With the higher molecular weight stuff
        our sections remain attached throughout in situ hybridizations, a rather        rigorous treatment for tissues.
B) You need to be sure that your slides are completely clean before poly-lysine
        treatment.  Packaged slides marked as "pre-cleaned" are no such thing!
        They have a thin coating of oil on them to prevent sticking together.
        I usually soak a box of slides in 70% ethanol + a drop or two of
        glacial acetic. I then remove the slides as needed and wipe them until
        dry with a Kimwipe.
Hope that this is helpful.
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Melissa Melan                           mmelan at lucy.wellelsey.edu
Department of Biological Sciences
Wellesley College
Wellelsey, MA  02181
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