BS³ (Bis[sulfosuccinimidyl] suberate)

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.

BS³ Highlights

• Amine-reactive Sulfo-NHS ester reacts rapidly with any primary amine-containing molecule
• BS³ is water-soluble, so crosslinking can be done in physiologic solutions
• Water-soluble crosslinking reagent is ideal for cell-surface crosslinking
• High-purity, crystalline BS³ can be used to create high-purity crosslinked conjugates

Applications for BS³

• 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

Product Details:

Properties of BS3
Alternate names	Sulfo-DSS
Molecular formula	C16H18N2O14S2Na2
Molecular weight	572.43
Spacer arm length	11.4 Å (8 atoms)
CAS Number	82436-77-9
Storage conditions	4°C, protect from moisture, use only fresh solutions
Reactive groups	NHS esters, react with primary amines at pH 7.0-9.0

BS³ Specifications

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

References:

1. Dihazi, G.H. and Sinz, A. (2003). Rapid Commun. Mass Spectrom. 17, 2005-2014.
2. Koller, D., et al. (2004) Selective Inactivation of Adrenomedullin over Calcitonin Gene-related Peptide Receptor Function by the Deletion of Amino Acids 14-20 of the Mouse Calcitonin-like Receptor. J. Biol. Chem. 279, 20387-20391.
3. Law, B.K., et al. (2002) Rapamycin Potentiates Transforming Growth Factor ß-Induced Growth Arrest in Nontransformed, Oncogene-Transformed, and Human Cancer Cells. Mol. Cell. Biol. 22, 8184-8198.
4. Mattson, G., et al. (1993). A practical approach to crosslinking. Molecular Biology Reports 17, 167-183.
5. Muller, D.R., et al. (2001). Anal. Chem. 73, 1927-1934.
6. Partis, M.D., et al. (1983). Crosslinking of proteins by omega-maleimido alkanoyl
   N-hydroxysuccinimide esters. J. Protein. Chem. 2, 263-277.
7. Sinz, A. (2003). J. Mass Spectrom. 38, 1225-1237.
8. Schutz, D.M., et al. (2004). Biochemistry 43, 4703-4715.

Related Products:

BS3 mass tag pairs
DSS
Bioconjugate Techniques (book)
EDC
Sulfo-SMCC

Related Resources:

Crosslinker Selection Guide