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Activated Carrier Proteins: Adar's Maleimide-Activated KLH
/Product Detail : Cat. 1027-Bulk Activated Carrier Proteins: Adar's Maleimide-Activated KLH
Product name : Activated Carrier Proteins: Adar's Maleimide-Activated KLH
Catalog number : Cat. 1027-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 these small molecules. KLH is
most popular for immunization while BSA conjugate is used in immunoassays of
the resulting antibodies in order to filter out the anti KLH response.
The Maleimide activated KLH and BSA produced by Adar Biotech are preactivated
with a heterobifunctional cross-linker (GMBS). These activated proteins
can be reacted with compounds that contain a free sulfhydryl group to form a stable
Maleimide-Activated KLH characteristics.
Activation method: GMBS.
Binding capacity: ~2-4 mg of peptide (average MW of 1000-2500) per 2 mg KLH
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 x1.0 to
x0.4 times the volume of Activated KLH solution. For example dissolve 2 mg of
peptide in 200-500 μl of buffer for addition to 2 mg of activated KLH in 500 μl.
Note: For haptens with limited solubility in H2O, DMSO may be used for its 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 KLH at room temperature.
Note: Do not vortex or heat the activated KLH.
3. Immediately mix the peptide and activated KLH and react for 2 hours at room
temperature under occasional mixing.
4. Peptide-conjugated KLH 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-KLH should be stored frozen until use.
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