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.
- Reactive groups: NHS ester (both ends)
- Reactive towards: amino groups (primary amines)
- Amine reactive sulfo-NHS ester reacts rapidly with any primary amine-containing molecule
- Membrane-permeable, allowing for intracellular crosslinking
- High purity, crystalline reagent can be used to create high-purity conjugates
- Water-insoluble (dissolve first in DMF or DMSO); compare to BS3 (Sulfo-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
|Chemical structure of DSS crosslinking reagent.
Properties of DSS.
|Spacer arm length
||11.4 Å (8 atoms)
||4°C, protect from moisture, use only fresh solutions
||NHS esters, react with primary amines at pH 7.0-9.0
We manufacture 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: > 9.2mg/mL in DMF and DMSO, clear and colorless solution
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- 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.
Crosslinker Application Guide -- search for recent literature references for this product
Overview of crosslinking
Chemistry of crosslinking
Crosslinkers at a glance (selection guide)
BS3 (water-soluble equivalent)
DSP (cleavable equivalent)
Bioconjugate Techniques (book)