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[Celegans] subcellular fractionation and mAbs

Morwenna Porter via celegans%40net.bio.net (by morwenna.porter from yale.edu)
Tue Apr 3 17:08:29 EST 2007


I'm doing some basic subcellular fractionation experiments in worms, separating
nuclei, membranes and cytosolic components, and to confirm I have done this
correctly I'm looking for a couple of nice monoclonal antibodies that
specifically recognize a transmembrane protein and a nuclear protein. Ideally
something that recognizes proteins of ~30-150kDa and which has been shown to
work well on worm westerns. Can anyone recommend anything?

Also, I'm doing these experiments by lysing worms in a French press, doing a
slow speed spin to remove carcasses etc and then a high speed spin at 100 000g
to separate soluble and insoluble fractions. However, this may not be the best
approach and I'm not quite sure where my nuclei would be? So if anyone has any
experience with these kinds of experiments and can offer some advice that would
be great too!

Thanks in advance,

Morwenna Porter, Ph.D
Postdoctoral Associate
Department of Molecular Biophysics and Biochemistry
Yale University School of Medicine
333 Cedar Street, SHM CE30
P.O. Box 208024
New Haven, CT 06520-8024

Tel: (203) 737-2271
Fax: (203) 785-6404
Email: morwenna.porter from yale.edu

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