Thermo Scientific Pierce DTSSP is a popular water-soluble crosslinker that contains amine-reactive NHS-ester ends around an 8-atom spacer arm, whose central disulfide bond can be cleaved with reducing agents.
DTSSP is 3,3´-Dithiobis(sulfosuccinimidylpropionate), which 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.
- Reactive groups: sulfo-NHS ester (both ends)
- Reactive towards: amino groups (primary amines)
- Sulfo-NHS ester reacts rapidly with any primary amine-containing molecule
- Disulfide bond in the spacer arm is readily cleaved by 10-50mM DTT or TCEP at pH 8.5
- Spacer arm also easily cleaved with reducing SDS-PAGE sample loading
- Cleavable crosslinker allows separation of crosslinked products
- Water-soluble; compare with DSP
- Membrane-impermeable, allowing for cell surface labeling
- 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
|Chemical structure of DTSSP crosslinking reagent.
Properties of DTSSP.
|Spacer arm length
||12.0 Å (8 atoms)
||4°C, protect from moisture, use only fresh solutions
||Sulfo-NHS esters, react with primary amines at pH 7.0-9.0
We manufacture 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.
- Solubility: >6mg/mL in DI water
- Identity: IR scan shows only peaks characteristic of the structure and functional groups of DTSSP
- Purity:> 80% by quantitative NMR (the highest standard for crosslinker purity)
- Back, J. W., et al. (2002). Protein Sci. 11, 2471-2478.
- Bennett, K.L., et al. (2000). Protein Sci. 9, 1503-1518.
- Cheng, S.K., et al. (2004) PloS Biology.2, e30.
- Paine-Saunders, S., et al. (2002). J. Biol. Chem.277, 2089-2096.
- Liliana Schaefer, L., et al. (2005). J. Clin. Invest.115, 2223-2233.
Crosslinker Application Guide -- search for recent literature references for this product
Overview of crosslinking
Chemistry of crosslinking
Crosslinkers at a glance (selection guide)
DSP (membrane permeable)
Bioconjugate Techniques (book)