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storage temp.
2-8°C
Specificity
KLH and BSA have been activated with succinimidyl-3-N-maleimidopropionic acid to provide a maleimide group at the end of a three atom spacer. Under approximately neutral conditions (pH 6.5-7.5) the maleimide will form a thioether bond with a sulfhydryl group, i.e., a cysteine residue on a peptide.
Features and Benefits
- Conjugation and purification in under 3 hours
- BSA and KLH are pre-activated and ready to use
- Mild reaction conditions: pH 6.5-7.5 in aqueous buffer
- Fast gel filtration separation of conjugate and free hapten
- Prepare immunogen and antigen for immunoassay with one kit
- Contains sufficient reagents for 3 conjugations each of BSA and KLH. Each reaction can conjugate at least 4 mg of hapten.
- Contains reagents for coupling efficiency assay
- Complete protocols and references included
Caution
Alkaline pH values (above 7.5) may hydrolyze the maleimide group or generate side reactions with amines. Haptens must contain cysteine or a sulfhydryl group in the reduced state in order to react efficiently with the maleimide group.
Principle
Sigma offers a kit for conjugating haptens to bovine serum albumin (BSA) and keyhole limpet hemocyanin (KLH). BSA and KLH have been used as protein carriers for haptens in immunization and immunoassay protocols. Immunogen is prepared with one carrier protein, typically KLH, while the antigen for immunoassay is prepared with another carrier, typically BSA. This eliminates the need to remove antibodies developed against the immunogen carrier protein from the antiserum before assay.
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D S Jones et al.
Bioconjugate chemistry, 10(3), 480-488 (1999-05-29)
Antibodies which bind beta2-glycoprotein I (beta2GPI) are associated with antiphospholipid syndrome. Synthetic peptide mimotopes have been discovered which compete with beta2GPI for binding to selected anti-beta2GPI. A thiol-containing linker was attached to the N-terminus of two cyclic thioether peptide mimotopes
B G Rose et al.
Bioconjugate chemistry, 6(5), 529-535 (1995-09-01)
Ceftiofur sodium is a broad spectrum, beta-lactamase-resistant cephalosporin. Ceftiofur and desfuroylceftiofur were used to develop competitive indirect enzyme-linked immunosorbent assays (CI-ELISA) for the determination of ceftiofur sodium. Hapten-protein conjugates were made using three different carrier proteins and three methods of
Tissue sulfhydryl groups.
G L ELLMAN
Archives of biochemistry and biophysics, 82(1), 70-77 (1959-05-01)
Kai Deng et al.
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Cross-linking of protein by ω-maleimido alkanoyl N-hydroxysuccinimido esters.
Partis, M. D., et al.
Journal of Protein Chemistry, 2, 263-277 (1983)
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