Thermo Scientific Pierce BS3 (Sulfo-DSS) is bis(sulfosuccinimidyl)suberate, an amine-to-amine crosslinker that is homobifunctional, water-soluble, non-cleavable and membrane impermeable.
BS3 contains an amine-reactive N-hydroxysulfosuccinimide (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-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 NHS-ester crosslinking reagents.
Because it contains the hydrophilic sulfonyl moiety, BS3 crosslinker is soluble up to ~100mM in water and many commonly used buffers, thus avoiding the use of organic solvents which may perturb protein structure. DSS, the non-water soluble analog of BS3 is also available for applications that require a less hydrophilic crosslinker (e.g., to effect intracellular crosslinking). DSS and BS3 have essentially identical crosslinking activity toward primary amines.
- Reactive groups: sulfo-NHS ester (both ends)
- Reactive towards: amino groups (primary amines)
- Amine-reactive Sulfo-NHS ester reacts rapidly with any primary amine-containing molecule
- Water-soluble; compare with DSS
- Membrane-impermeable, allowing for cell surface labeling
- High-purity, crystalline reagent can be used to create high-purity crosslinked conjugates
- Crosslink cell-surface proteins prior to cell lysis and immunoprecipitation
- Identify receptor-ligand interactions by chemical crosslinking
- “Fix” protein interactions to allow identification of weak or transient protein interactions
- Identify near-neighbor protein interactions
- Protein crosslinking to create bioconjugates via single-step reactions
- Immobilize proteins onto amine-coated surfaces
|Chemical structure of BS3 crosslinking reagent.
Properties of BS3.
|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 BS3 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 BS3 is tested to meet the following minimum specifications.
- Purity: > 93% by quantitative NMR (the highest standard for crosslinker purity)
- Solubility: > 5.8 mg/mL in DI water, clear solution with no insoluble material
- Dihazi, G.H. and Sinz, A. (2003). Rapid Commun. Mass Spectrom. 17, 2005-2014.
- Koller, D., et al. (2004). J. Biol. Chem. 279, 20387-20391.
- Law, B.K., et al. (2002). Mol. Cell. Biol. 22, 8184-8198.
- Mattson, G., et al. (1993). A practical approach to crosslinking. Molecular Biology Reports 17, 167-183.
- Muller, D.R., et al. (2001). Anal. Chem. 73, 1927-1934.
- Partis, M.D., et al. (1983). Crosslinking of proteins by omega-maleimido alkanoyl
N-hydroxysuccinimide esters. J. Protein. Chem. 2, 263-277.
- Sinz, A. (2003). J. Mass Spectrom. 38, 1225-1237.
- Schutz, D.M., et al. (2004). Biochemistry 43, 4703-4715.
Crosslinker Application Guide -- search for recent literature references for this product
Overview of crosslinking
Chemistry of crosslinking
Crosslinkers at a glance (selection guide)
BS3 mass tag pairs
DSS (membrane-permeable equivalent)
DTSSP (cleavable equivalent)
Bioconjugate Techniques (book)