MBK1
Sigma
Maleimide Activated BSA, KLH Conjugation Kit
Description
| 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. |
| 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 |
| 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. |
| 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. |
Properties
| storage temp. | 2-8°C |
Safety
Related Products
References
| reference | Jones, D.S., et al., Multivalent thioether-peptide conjugates: B cell tolerance of an anti-peptide immune response. Bioconjug. Chem. 10, 480-488, (1999) Abstract |
| Rose, B.G., et al., Competitive indirect ELISA for ceftiofur sodium and the effect of different immunizing and coating antigen conjugates. Bioconjug. Chem. 6, 529-535, (1995) Abstract | |
| Partis, M. D., et al., Cross-linking of protein by ω-maleimido alkanoyl N-hydroxysuccinimido esters. J. Protein Chem. 2, 263-277, (1983) | |
| Ellman, G.L., Tissue sulfhydryl groups. Arch. Biochem. Biophys. 82, 70-77, (1959) Abstract |
