Thermo Scientific Pierce Sulfo-SMCC is a very popular, water-soluble, amine-to-sulfhydryl crosslinker that contains NHS-ester and maleimide reactive groups at opposite ends of a medium-length cyclohexane spacer arm (8.3 angstroms).
Sulfosuccinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (Sulfo-SMCC) is a water-soluble, non-cleavable and membrane impermeable crosslinker. It contains an amine-reactive N-hydroxysuccinimide (NHS ester) and a sulfhydryl-reactive maleimide group. NHS esters react with primary amines at pH 7-9 to form stable amide bonds. Maleimides react with sulfhydryl groups at pH 6.5-7.5 to form stable thioether bonds. The maleimide groups of Sulfo-SMCC and SMCC are unusually stable up to pH 7.5 because of the cyclohexane bridge in the spacer arm. Because it contains the hydrophilic sulfonyl moiety, Sulfo-SMCC is soluble up to ~10 mM in water and many commonly used buffers, thus avoiding the use of organic solvents which may perturb protein structure.
Sulfo-SMCC Highlights
- Amine reactive Sulfo-NHS ester crosslinks rapidly with primary amine-containing molecule
- Sulfhydryl-reactive maleimide reacts with cysteine residues to yield specific conjugates
- Sulfo-SMCC is water soluble, so crosslinking can be done in physiologic solutions
- High purity, crystalline Sulfo-SMCC can be used to create high-purity maleimide-activated derivatives
- Cyclohexane bridge confers added stability to the maleimide group making Sulfo-SMCC the ideal crosslinking agent for maleimide activation of proteins. Maleimide groups are stable for 64 hours in 0.1 M sodium phosphate buffer, pH 7 at 4°C.
Applications for Sulfo-SMCC
- Ideal crosslinker for enzyme labeling of antibodies, both enzyme activity and antibody specificity can be preserved
- Create specific bioconjugates via one- or two-step crosslinking reactions
- Create sulfhydryl-reactive, maleimide-activated carrier proteins for coupling haptens
Product Details:
Sulfo-SMCC is often used for the preparation of antibody-enzyme and hapten-carrier conjugates. In this type of conjugation, the NHS ester is reacted first with the antibody, excess crosslinking reagent removed and then the sulfhydryl-containing enzyme molecule is added. This two-step reaction scheme results in formation of specific antibody-enzyme conjugates.
Sulfo-SMCC is the reagent of choice for creating stable maleimide-activated carrier proteins that will spontaneously react with sulfhydryls. In this application, the NHS ester of Sulfo-SMCC is reacted with lysine residues on the carrier protein, converting them to reactive maleimides. These relatively stable maleimide-activated intermediates may be lyophilized and stored for later conjugation to a hapten.
| Properties of Sulfo-SMCC |
| Molecular formula |
C16H17N2O9SNa |
| Molecular weight |
436.37 |
| Spacer arm length |
8.3 Å (9 atom) |
| CAS Number |
92921-24-9 |
| Storage conditions |
-20°C, protect from moisture, use only fresh solutions |
| Reactive groups: |
NHS ester, reacts with primary amines at pH 7.0-9.0 |
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Maleimide, reacts with sulfhydryls at pH 6.5-7.5 |
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Sulfo-SMCC Specifications. We manufacture Sulfo-SMCC to the highest specifications to produce the most specific bioconjugates and maleimide-activated proteins, ensure the integrity of your data and to provide you with the highest degree of crosslinking consistency. Each lot of Sulfo-SMCC is tested to meet the following minimum specifications.
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Identity
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IR scan shows only peaks characteristic of the structure and functional groups of Sulfo-SMCC crosslinker |
| Purity |
≥ 90% by quantitative NMR (the highest standard for crosslinker purity) |
| Solubility |
≥ 10 mg/ml in DI water |
References:
- Muller L. et al. (2010) Evolutionary gain of function for the ER membrane protein Sec62 from yeast to humans. Mol. Biol. Cell. 21, 691-703.
- Dickgreber N. et al. (2009) Targeting antigen to MHC class II molecules promotes efficient cross-presentation and enhances immunotherapy. J. Immunol. 182, 1260-9.
- Kersemans V. et al. (2008) Drug-resistant aml cells and primary aml specimens are killed by 111In-anti-CD33 monoclonal antibodies modified with nuclear localizing peptide sequences. J. Nucl. Med. 49, 1546-54.
- Duval M. et al. (2008) A bispecific antibody composed of a nonneutralizing antibody to the gp41 immunodominant region and an anti-CD89 antibody directs broad human immunodeficiency virus destruction by neutrophils. J. Virol. 82, 4671-4.
- Costantini D. L. et al. (2008) Trastuzumab-resistant breast cancer cells remain sensitive to the auger electron-emitting radiotherapeutic agent 111In-NLS-Trastuzumab and are radiosensitized by methotrexate. J. Nucl. Med. 49, 1498-505.
- Brinkmann M. M. et al. (2007) The interaction between the ER membrane protein UNC93b and TLR3, 7, and 9 is crucial for TLR signaling. J. Cell Biol. 177, 265-75.
- Kehoe J. W. et al. (2006) Using phage display to select antibodies recognizing post-translational modifications independently of sequence context. Mol. Cell. Proteomics. 5, 2350-63.
Related Products:
SMCC
Bioconjugate Techniques (book)
Maleimide Activated Carrier Proteins and Kits
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