3,3´-Dithiobis(sulfosuccinimidylpropionate)  (DTSSP) is a water-soluble, homobifunctional, thiol-cleavable and membrane impermeable crosslinker. It contains an amine-reactive N-hydroxysulfosuccinimide (sulfo-NHS) ester at each end of an 8-carbon spacer arm. Sulfo-NHS esters react with primary amines at pH 7-9 to form stable amide bonds, along with release of the N-hydroxysulfosuccinimide 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 sulfo-NHS-ester crosslinking reagents. DSS, the non-sulfonated analog of DTSSP is also available for applications that require a membrane-permeable crosslinker. BS³, the non-cleavable analog of DTSSP is also available for applications that require a stable spacer arm that cannot be cleaved in the presence of reducing agents.

DTSSP Highlights

• Amine reactive sulfo-NHS ester reacts rapidly with any primary amine-containing molecule
• Disulfide bond in the spacer arm is readily cleaved by 10-50 mM DTT or TCEP at pH 8.5
• Spacer arm is also cleaved with 5% β-mercaptoethanol in SDS-PAGE sample loading buffer at 100°C for 5 minutes
• Cleavable crosslinker allows separation of crosslinked products
• DTSSP crosslinker is not membrane permeable, so cell-surface crosslinking can be done

Applications for DTSSP

• Chemical crosslinking of cell surface 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

DTSSP Specifications
Pierce manufactures DTSSP 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 DTSSP is tested to meet the following minimum specifications.

Related Products
BS³         DSP
Bioconjugate Techniques

Guide: Crosslinker Selection Guide
Tech Tip: Determine reactivity of NHS ester biotinylation and crosslinking reagents

References

1. Back, J. W., et al. (2002). Protein Sci. 11, 2471-2478.
2. Bennett, K.L., et al. (2000). Protein Sci. 9, 1503-1518.
3. Cheng, S.K., et al. (2004) PloS Biology.2, e30.
4. Paine-Saunders, S., et al. (2002) Heparan Sulfate Proteoglycans Retain Noggin at the Cell Surface. A Potential Mechanism for Shaping Bone Morphogenetic Protein Gradients.J. Biol. Chem.277, 2089-2096.
5. Liliana Schaefer, L., et al. (2005) The matrix component biglycan is proinflammatory and signals through Toll-like receptors 4 and 2 in macrophages. J. Clin. Invest.115, 2223-2233.

* DTSSP is available in bulk quantities for manufacturing applications.