DSS (disuccinimidyl suberate), No-Weigh™ Format - FAQs

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10 product FAQs found

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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Is the DSS crosslinking reagent water-soluble?

No. The reagent must first be reconstituted in an organic solvent before 10-fold dilution in aqueous buffers. The organic solvent should not exceed 10% of the total react volume.

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Is a hazy solution common after addition of the dissolved DSS solution to an aqueous buffer solution?

Yes. The resultant reaction mixture will become hazy frequently after the addition of the reagent solution.

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How much DSS crosslinking reagent should I use?

We typically use a 10-fold molar excess of crosslinker to protein for solutions greater than 5 mg/mL. Use a 20- to 50-fold molar excess for more dilute protein solutions.

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How is the DSS crosslinking reaction quenched?

The reaction is quenched by the addition of 1 M Tris or glycine, pH 7.5 to the reaction mixture to a final concentration of 20-50 mM.

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Can I make a stock solution of the DSS crosslinker for long-term storage?

Stock solutions for long-term storage are NOT recommended because NHS esters are extremely vulnerable to hydrolysis. Even organic solvents are hygroscopic and tend to absorb water and promote hydrolysis. The crosslinker should be made immediately before each use.

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