crosslinking reagents for nuclear protein complexes
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.
Zhonglin Chai, PhD
Baker Medical Research Institute, Australia
>>> Britney <I'mNotReallyBritneySoPleaseDon'tEmailMe at hotmail.com> 01/16/01 04:53pm >>>
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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