Thermo Scientific Pierce LC-SMCC is an amine-to-sulfhydryl crosslinker that contains NHS-ester and maleimide reactive groups at opposite ends of a long cyclohexane-stabilized spacer arm (16.2 angstroms).
LC-SMCC is a non-cleavable and membrane permeable 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 LC-SMCC and are unusually stable up to pH 7.5 because of the cyclohexane bridge in the spacer arm.
LC-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.
LC-SMCC is a reagent of choice for creating stable maleimide-activated carrier proteins that will spontaneously react with sulfhydryls. In this application, the NHS ester of LC-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.
Highlights:
- Amine reactive NHS ester crosslinks rapidly with primary amine-containing molecule
- Sulfhydryl-reactive maleimide reacts with cysteine residues to yield specific conjugates
- High purity, crystalline LC-SMCC can be used to create high-purity maleimide-activated derivatives
- Cyclohexane bridge confers added stability to the maleimide group making LC-SMCC an 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:
- Ideal reagent 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:
| Properties of LC-SMCC. |
| Molecular formula |
C22H29N3O7 |
| Molecular weight |
447.48 |
| Spacer arm length |
16.1Å (16 atoms) |
| 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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LC-SMCC Specifications. We manufacture LC-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 LC-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 LC-SMCC cross linker |
| Purity |
≥ 90% by NMR |
References:
- Bieniarz, C., et al. (1996). Extended length Heterobifunctional Coupling Agents for Protein Conjugations. Bioconjug. Chem. 7, 88-95.
- Kuijpers, W.H., et al. (1993). Bioconjug. Chem. 4(1), 94-102.
- Mattson, G., et al. (1993). A practical approach to crosslinking. Molecular Biology Reports 17, 167-183.
- Brinkley, M.A. (1992). A survey of methods for preparing protein conjugates with dyes, haptens and crosslinking reagents. Bioconjugate Chem. 3, 2-13.
- Uto, I., Ishimatsu, T., Hirayama, H., Ueda, S., Tsuruta, J. and Kambara, T. (1991). Determination of urinary Tamm-Horsfall protein by ELISA using a maleimide method for enzyme-antibody conjugation. J. Immunol. Methods 138, 87-94.
- Partis, M.D., et al. (1983). Crosslinking of proteins by omega-maleimido alkanoyl N-hydroxysuccinimide esters. J. Protein. Chem. 2, 263-277.
- Yoshitake, S., et al. (1982). Mild and efficient conjugation of rabbit Fab and horseradish peroxidase using a maleimide compound and its use for enzyme immunoassay. J. Biochem. 92, 1413-1424.
Related Products:
SMCC
Sulfo-SMCC
Bioconjugate Techniques (book)
Maleimide Activated Carrier Proteins and Kits
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