Thermo Scientific Pierce DSS, also called disuccinimidyl suberate, is noncleavable and membrane permeable crosslinker that contains an amine-reactive N-hydroxysuccinimide (NHS) ester at each end of an 8-carbon spacer arm.
NHS esters react with primary amines at pH 7-9 to form stable amide bonds, along with release of the N-hydroxysuccinimide leaving group. Proteins, including antibodies, generally have several primary amines in the side chain of lysine (K) residues and the N-terminus of each polypeptide that are available as targets for NHS-ester crosslinking reagents.
DSS is first dissolved in an organic solvent such as DMF or DMSO, then added to the aqueous crosslinking reaction. BS3, the water soluble analog of DSS is also available for applications that require a hydrophilic crosslinker (e.g. to effect cell-surface crosslinking). DSS and BS3 have essentially identical crosslinking activity toward primary amines.
DSS Highlights
- Amine reactive Sulfo-NHS ester reacts rapidly with any primary amine-containing molecule
- DSS crosslinker is membrane permeable, so intracellular crosslinking can be done
- High purity, crystalline DSS can be used to create high-purity conjugates
Applications for DSS
- Chemical crosslinking of intracellular proteins prior to cell lysis and immunoprecipitation
- “Fix” protein interactions to allow identification of weak or transient protein interactions
- Protein crosslinking to create bioconjugates via single-step reactions
- Immobilize proteins onto amine-coated surfaces
Product Details:
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Properties of DSS
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Molecular formula
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C16H20N2O8 |
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Molecular weight
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368.35 |
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Spacer arm length
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11.4 Å (8 atoms) |
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CAS Number
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68528-80-3 |
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Storage conditions
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4°C, protect from moisture, use only fresh solutions |
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Reactive groups
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NHS esters, react with primary amines at pH 7.0-9.0 |
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DSS Specifications
Pierce manufactures DSS to the highest specifications to produce the most specific bioconjugates, ensure the integrity of your data and to provide you with the highest degree of consistency. Each lot of DSS crosslinker is tested to meet the following minimum specifications.
Identity IR scan shows only peaks characteristic of the structure and functional groups of DSS
Purity > 90% by quantitative NMR (the highest standard in crosslinking purity)
Solubility > 9.2 mg/ml in DMF and DMSO, clear and colorless solution
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References
- Chataway, T.K. and Barritt, G.J. (1995). Mol. Cell. Biochem. 145(2), 111-120.
- Donato, R., et al. (1989). J. Neurochem. 53(2), 566-571.
- Ishmael, F.T., et al. (2005) Intersubunit and Domain Interactions of the Meprin B Metalloproteinase: Disulfide Bonds and Protein-Protein Interactions in the MAM and TRAF Domains. J. Biol. Chem.280, 13895-13901.
- Longshaw, V.M., et al. (2004) Nuclear translocation of the Hsp70/Hsp90 organizing protein mSTI1 is regulated by cell cycle kinases. J. Cell Sci. 117, 701-710.
- Mattson, G., et al. (1993). A practical approach to crosslinking. Molecular Biology Reports 17, 167-183.
- Mathieu, M., et al. (1990). Mol. Endocrinol. 4(9), 1327-1335.
- Partis, M.D., et al. (1983). Crosslinking of proteins by omega-maleimido alkanoyl N-hydroxysuccinimide esters. J. Protein. Chem. 2, 263-277.
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