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Activated Carrier Proteins: Adar's Maleimide-Activated BSA
/Product Detail : Cat. 1025-Bulk Activated Carrier Proteins: Adar's Maleimide-Activated BSA
Product name : Activated Carrier Proteins: Adar's Maleimide-Activated BSA
Catalog number : Cat. 1025-Bulk
Supplier name : Adar
Data sheet : Ask more or other datasheet now !
Activated KLH and BSA are commonly used as protein carriers for haptens such as
peptides in order to enable the immune response to small molecules. In many cases
KLH conjugate is used for the immunization, while BSA conjugate is used in
immunoassys of the resulting antibodies. BSA conjugates are used in order to filter
out the antibodies directed to the KLH. However in some cases BSA or other
proteins are preffered as carrier proteins because of a variety of other reasons.
The Maleimide activated KLH and BSA produced by Adar Biotech are
preactivated with a heterobifunctional cross-linker (GMBS). These activated
proteins may be reacted with biomolecules that contain a free sulfhydryl groups, to
form a stable thioether bond.
Maleimide-Activated BSA characteristics.
Activation method: GMBS.
Binding capacity: ~2.5-4 mg of peptide (average MW of 1000-2500) per 2 mg BSA
Protein concentration: 4 mg/ml (0.5 ml)
Storage buffer: PBS pH 7.5
Storage condition: -20°C.
A. Procedure for Peptide Conjugation
1. Dissolve the sulfhydryl-containing hapten in a volume of water equal to 1.0-2.5
times the volume of BSA. For example dissolve 2 mg of peptide in 200-500 μl of
buffer for addition to 2 mg of activated BSA in 500 μl.
Note: For haptens with limited solubility, DMSO may be used for solubilization. Use .30%
DMSO in the final conjugation solution or the carrier protein may irreversibly denature.
Alkaline pH values (above 8.5) 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.
2. Thaw the Maleimide Activated BSA at room temperature.
Note: Do not vortex or heat the activated BSA.
3. Immediately mix the peptide and activated BSA and react for 2 hours at room
4. Peptide-conjugated BSA can be purified by gel filtration or dialysis to remove
Note: If the immunogen is to be stored for more than a few days it is recommended to store
frozen at -20°C.
5. The coupling efficiency of conjugation can be determined by assaying the
content of free sulfhydryl groups in the unreacted peptide using DTNB reagent.
The activated-BSA should be stored frozen until use.
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