Will the DSS crosslinked product become unlinked as a result of boiling, incubation with SDS, high salt, high or low pH or any other means?
No. DSS crosslinks are amide bonds (same as peptide bonds).
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What is the recommended buffer for optimal DSS crosslinking?
We recommend the following buffers: 20 mM sodium phosphate, 0.15 M NaCl, PBS, pH 7.5. Or HEPES, bicarbonate-carbonate and borate buffers may be substituted for PBS. Nearly any non-amine buffer pH 7-8 can be used.
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What is the difference between DSS and BS3 crosslinkers?
BS3 is the water-soluble analogue of DSS (i.e., DSS is the membrane-permeable analog of BS3). DSS is water-insoluble and membrane-permeable, so it can be used for both intracellular and cell surface crosslinking. BS3 is water-soluble and membrane-impermeable, so it can be used for cell-surface crosslinking.
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What buffers are not recommended for DSS crosslinking?
Any buffer containing primary amines (Tris, glycine, lysine, and ethanolamine) will not work. Glutathione- and imidazole-containing buffers are also not recommended.
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What are the reactive groups in the DSS crosslinking reagent and what are they reactive toward?
The reactive groups are NHS esters that are reactive toward primary amines.
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