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crosslinking reagents for nuclear protein complexes

Zhonglin Chai Zhonglin.Chai at baker.edu.au
Tue Jan 16 01:11:38 EST 2001

DSP is water soluble and membrane permeable with a 12 A arm space and a disulfate bond that is cleavable under reducing conditions. It should be suitable for your purpose. It was also used to cross-link proteins in vivo and the treated cells remained viable. Probably you may want to add DSP directly to the cells and hope it getting into the nucleus and cross-linking the nuclear protein complex followed by cell lysis and IP.  I have not used BASED and have no words about it.

It would be interesting to hear how it goes with nuclear proteins.

Good Luck

Zhonglin Chai, PhD
Baker Medical Research Institute, Australia

>>> Britney <I'mNotReallyBritneySoPleaseDon'tEmailMe at hotmail.com> 01/16/01 04:53pm >>>
Dear All,
I am interested in analysing the protein constituents of a large protein
complex within the nucleas. I want to try taking a nuclear extract,
adding a crosslinker, immunoaffinity purifying the complex, reducing the
crosslinker, running the proteins on 1D- and/or 2D PAGE gels, then use
Mass spectrometry to identify the proteins. There are two protein
crosslinkers I want to purchase: DSP (Lomants reagent) and BASED. Before
I make the purchase, can anyone offer any comments or suggestions on
these or alternative crosslinkers?



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