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Maleimide Activated Horseradish Peroxidase and Kit


Conjugate HRP to sulfhydryl-containing molecular probes.

Pierce Maleimide Activated Horseradish Peroxidase

Thermo Scientific Pierce Maleimide Activated Horseradish Peroxidase and Kit is for preparation of HRP conjugates with proteins, peptides or other ligands that contain sulfhydryl groups, such as reduced cysteines.

This product consists of horseradish peroxidase (HRP) that has been modified with Sulfo-SMCC (Part No. 22322) to attach several maleimide groups per HRP molecule while retaining the peroxidase activity. The activated HRP will covalently attach to proteins or other molecule containing sulfhydryl groups (e.g., cysteines). HRP-conjugates of antibodies, proteins, peptides and other thiol-containing reporter probes are easily made using this method. The complete kit includes the activated HRP as well as two types of reagents for preparing sulfhydryl-ready antibodies (IgG) or proteins for conjugation.

Highlights:

  • Activated HRP – horseradish peroxidase (HRP) modified with maleimide groups for conjugation to sulfhydryl molecules
  • Sulfhydryl-reactivemaleimide groups conjugate to reduced thiols (-SH), as in the side-chain of cysteine residues
  • High activity HRP – enzyme activity is greater than 240 units/mg; lyophilized, activated enzyme is stable for at least 12 months at 4°C
  • Complete kit – includes the activated HRP as well as two types of reagents for sulfhydryl-ready antibodies (IgG) or proteins for conjugation

Product Details:

Sulfhydryl reaction scheme for maleimide crosslinkers, biotinylation reagents or labeling reagents.
Reaction scheme for conjugation of Maleimide Activated HRP to a sulfhydryl. In this diagram, (R) represents the maleimide-activated peroxidase; (P) represents a protein or other molecule that contains the target functional group (i.e., sulfhydryl, –SH). For more information, see our review of Maleimide Reaction Chemistry.

The complete kit for Maleimide Activated Horseradish Peroxidase contains reagents for exposing or added the necessary sulfhydryl groups on antibodies (IgG) or practically any other protein. These general strategies are described briefly in the applications section of our review of Maleimide Reaction Chemistry. Of course, any protein that contains cysteines has sulfhydryl groups (-SH), but they must be reduced (not in the form of disulfide bonds) to be conjugated. Antibodies also contain disulfide bonds that can be targeted as antibody labeling sites; the hinge-region disulfide bonds in IgG can be selectively cleaved with the mild reducing agent 2-Mercaptoethylamine (Part No. 20408), which is included in the complete kit. Alternatively, sulfhydryl groups can be added to proteins (or any amine-containing molecule) using SATA reagent (Part No. 26102), which also is included in the kit.

References:

  1. Hashida, S., et al. (1984). More useful maleimide compounds for the conjugation of Fab' to horseradish peroxidase through thiol groups in the hinge. J. Appl. Biochem. 6, 56-63.
  2. Imagawa, M., et al.(1982). Characteristics and evaluation of antibody-horseradish peroxidase conjugates prepared by using a maleimide compound, glutaraldehyde, and periodate. J. Appl. Biochem. 4, 41-57.
  3. O’Sullivan, M.J., et al. (1979). Comparison of two methods of preparing enzyme-antibody conjugates: application of these conjugates for enzyme immunoassay. Anal. Biochem. 100, 100-108.

Related Resources:

Immunohistochemistry (IHC) technical guide
Immunodetection for IHC technical guide

Related Products:

Plus Activated Peroxidase Kit
Anti-HRP Monoclonal Antibody (Part No. MA1-10371)


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
31485 EZ-Link Maleimide Activated Horseradish Peroxidase
Formulation: Activated protein lyophilized from PBS
Sufficient For: Conjugation with approx. 5mg of antibody (IgG)
5mg Product Instructions for product #31485 EZ-Link Maleimide Activated Horseradish Peroxidase MSDS for product #31485 EZ-Link Maleimide Activated Horseradish Peroxidase Certificate of Analysis for product #31485 EZ-Link Maleimide Activated Horseradish Peroxidase $198.00

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31485B EZ-Link Maleimide Activated Horseradish Peroxidase
Formulation: Activated protein lyophilized from PBS
Custom MSDS for product #31485B EZ-Link Maleimide Activated Horseradish Peroxidase Certificate of Analysis for product #31485B EZ-Link Maleimide Activated Horseradish Peroxidase
31494 EZ-Link Maleimide Activated Horseradish Peroxidase Kit
Formulation: Multi-component kit
Sufficient For: 1 enzyme-labeling reaction with up to 5mg of antibody (IgG)

Kit Contents:
Maleimide Activated Horseradish Peroxidase, 5mg
Conjugation Buffer (10X), 20mL
PBS Pack (makes 500mL), 1 pack
2-Mercaptoethylamine-HCl, 6mg
SATA, 2mg
Dimethylformamide, 1mL
Hydroxylamine-HCl, 5mg
Polyacrylamide Desalting Column, 10mL, 1 column
1-rxn kit Product Instructions for product #31494 EZ-Link Maleimide Activated Horseradish Peroxidase Kit MSDS for product #31494 EZ-Link Maleimide Activated Horseradish Peroxidase Kit Certificate of Analysis for product #31494 EZ-Link Maleimide Activated Horseradish Peroxidase Kit $313.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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