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.
- Reactive groups: sulfo-NHS ester and maleimide
- Reactive towards: amino and sulfhydryl groups
- Water-soluble (compare to SMCC), so crosslinking can be done in physiologic solutions
- High purity, crystalline reagent 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.
- Enzyme labeling of antibodies - both enzyme 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
|Chemical structure of Sulfo-SMCC crosslinking reagent.
|Antibody-enzyme conjugation with Sulfo-SMCC. This conjugation method enables preparation of maleimide-activated protein (step 1), which can be stably stored for later conjugation to a sulfhydryl-containing protein (step 2). Depending upon the availability of amines and sulfhydryls on respective proteins, the approach can be used in either direction (e.g., enzyme-to-antibody or antibody-to-enzyme). A very popular application for Sulfo-SMCC is activation of immunogenic carrier proteins for conjugation to cysteine-terminated peptides for use in antibody production (e.g., Part No. 77606).
Properties of Sulfo-SMCC.
|Spacer arm length
||8.3 Å (9 atom)
||-20°C, protect from moisture, use only fresh solutions
||Sulfo-NHS ester, reacts with primary amines at pH 7.0-9.0
||Maleimide, reacts with sulfhydryls at pH 6.5-7.5
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.
- Identity: 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: ≥ 10mg/mL in DI water
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- 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.
Crosslinker Application Guide -- search for recent literature references for this product
Overview of crosslinking
Chemistry of crosslinking
Crosslinkers at a glance (selection guide)
Bioconjugate Techniques (book)
Maleimide Activated Carrier Proteins and Kits