Thermo Scientific Pierce DSP (Lomant's Reagent) is an amine-reactive crosslinker having NHS-ester reactive ends and a cleavable disulfide bond in its spacer arm.
DSP 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-hydroxy-succinimide 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, the non-cleavable analog of DSP crosslinker is also available for applications that require a stable spacer arm that cannot be cleaved in the presence of reducing agents.
- Reactive groups: NHS ester (both ends)
- Reactive towards: amino groups (primary amines)
- Amine-reactive 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 cleavable with reducing SDS-PAGE sample loading buffer
- Cleavable crosslinker allows separation of crosslinked products
- Water-insoluble (dissolve first in DMF or DMSO); compare to DTSSP (Sulfo-DSP)
- Membrane-permeable, allowing for intracellular crosslinking
- Chemical crosslinking of intracellular proteins prior to cell lysis and immunoprecipitation
- “Fix” protein interactions to allow identification of weak or transient protein interactions
- Fix tissues for immunostaining without damaging RNA
- Protein crosslinking to create bioconjugates via single-step reactions
- Immobilize proteins onto amine-coated surfaces
|Chemical structure of DSP crosslinking reagent.
Properties of DSP.
||Lomant’s reagent, DTSP
|Spacer arm length
||12.0 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 DSP 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 DSP is tested to meet the following minimum specifications.
- Identity: IR scan shows only peaks characteristic of the structure and functional groups of DSP
- Purity: > 95% by quantitative NMR (the highest standard for crosslinker purity)
- Adir, N. and Ohad, I. (1986). Biochim. Biophys. Acta 850(2), 264-274.
- Appenzeller-Herzog, C., et al. (2005) Carbohydrate- and Conformation-dependent Cargo Capture for ER-Exit Mol. Biol. Cell. 16, 1258-1267.
- Joshi, S. and Burrows, R. (1990). AT synthase complex from bovine heart mitochondria. J. Biol. Chem. 265, 14518-14525.
- Leduc, M., et al. (1989). Microbiol. Lett. 51(1), 11-14.
- Mattson, G., et al. (1993). A practical approach to crosslinking. Molecular Biology Reports 17, 167-183.
- Partis, M.D., et al. (1983). Crosslinking of proteins by omega-maleimido alkanoyl
N-hydroxysuccinimide esters. J. Protein. Chem. 2, 263-277.
- Xiang, C.C., et al. (2004) Using DSP, a reversible crosslinker, to fix tissue sections for immunostaining, microdissection and expression profiling. Nucleic Acids Res.32, e185.
Crosslinker Application Guide -- search for recent literature references for this product
Overview of crosslinking
Chemistry of crosslinking
Crosslinkers at a glance (selection guide)
DSS (non-cleavable equivalent)
DTSSP (water-soluble equivalent)
Bioconjugate Techniques (book)