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Index / Adar / Activated Carrier Proteins: Adar's Maleimide-Activated BSA /Product Detail : Cat. 1025-Bulk Activated Carrier Proteins: Adar's Maleimide-Activated BSA
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Activated Carrier Proteins: Adar's Maleimide-Activated BSA

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Product name : Activated Carrier Proteins: Adar's Maleimide-Activated BSA

Catalog number : Cat. 1025-Bulk

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Supplier name : Adar

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Introduction

 

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

temperature.

 

4. Peptide-conjugated BSA can be purified by gel filtration or dialysis to remove

unbound peptide 

 

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.  

 

B.  Storage

The activated-BSA should be stored frozen until use.

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