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Index / Adar / Activated Carrier Proteins: Adar's Maleimide-Activated KLH /Product Detail : Cat. 1027-10 (10mg) Activated Carrier Proteins: Adar's Maleimide-Activated KLH
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#Cat. 1027-10 (10mg)

Activated Carrier Proteins: Adar's Maleimide-Activated KLH

 Price: 353   EUR
400   USD
273   GBP

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

Catalog number : Cat. 1027-10 (10mg)

Quantity:

Availability: Yes

Supplier name : Adar

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More Details about

Introduction

 

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

thioether bond.  

 

 

 

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

excess 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-KLH should be stored frozen until use.

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