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P1709

Sigma-Aldrich

Peroxidase, Maleimide Activated from horseradish

lyophilized powder, >200 units/mg protein

Synonym(s):

Maleimide activated HRP

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About This Item

MDL number:
MFCD00284131
UNSPSC Code:
12352204
NACRES:
NA.54

biological source

horseradish

Quality Level

200

form

lyophilized powder

specific activity

>200 units/mg protein

composition

Protein, ~50% E1%/403

extent of labeling

1-3 mol maleimide per mol protein

storage temp.

−20°C

Related Categories

General description

Peroxidase (POD) is an enzyme that includes a group of specific enzymes such as nicotinamide adenine dinucleotide (NAD) peroxidase, NADP peroxidase, fatty acid peroxidase and a group of non-specific enzymes from various sources. It is present in animals, higher plants and other organisms.

Application

For direct conjugation to compounds containing free sulfhydryl groups for use as an enzyme-linked detection reagent.
Instructions for use included. In a study on horseradish peroxidase, the Maleimide compounds succinimidyl 4-maleimidobutyrate and succinimidyl 6-maleimidohexanoate have been shown to be the most effective in the conjugation of Fab′ to horseradish peroxidase. This enzyme has also been used in a study to investigate the effectiveness in staining a mitochondrial protein, lipoamide dehydrogenase.
To increase signal intensity of immunoconjugates, immunoglobulins were synthetically modified to contain multiple primary amines and covalently coupled to maleimide activated peroxidase. This can result in more efficient conjugation and signal amplification.

Biochem/physiol Actions

Peroxidase (POD) helps in the dehydrogenation of several organic compounds like phenols, aromatic amines, hydroquinones etc.

Other Notes

Type VI peroxidase activated with maleimidocaproic acid N-hydroxysuccinimide ester.

Unit Definition

One unit will form 1.0 mg of purpurogallin from pyrogallol in 20 seconds at pH 6.0 at 20°C.

Physical form

Lyophilized powder containing trehalose and sodium citrate

Pictograms

Health hazard
GHS08

Signal Word

Danger

Hazard Statements

H317 - H334

Hazard Classifications

Resp. Sens. 1 - Skin Sens. 1

Storage Class Code

11 - Combustible Solids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Certificates of Analysis (COA)

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Biochemical Methods (1996)
Subhash Dhawan
Peptides, 23(12), 2091-2098 (2003-01-22)
Immunoconjugates are widely used for indirect detection of analytes (such as antibodies or antigens) in a variety of immunoassays. However, the availability of functional groups such as primary amines or free sulfhydryls in an immunoglobulin molecule is the limiting factor
S Matuda et al.
In vivo (Athens, Greece), 1(4), 221-227 (1987-07-01)
IgG-maleimide peroxidase, Fab'-maleimide peroxidase, polymer and monomer types of Fab'-periodate peroxidase were prepared from an antibody against rat lipoamide dehydrogenase, a component of the pyruvate dehydrogenase complex which is located in mitochondria. They were examined for immunohistochemical staining of the
S Hashida et al.
Journal of applied biochemistry, 6(1-2), 56-63 (1984-02-01)
Nine different maleimide compounds were evaluated for the conjugation of Fab' to horseradish peroxidase through thiol groups in the hinge. The compounds evaluated were succinimidyl maleimidoacetate (I), succinimidyl 4-maleimidobutyrate (II), succinimidyl 6-maleimidohexanoate (III), succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (IV), succinimidyl m-maleimidobenzoate (V), succinimidyl
Kitagawa, T., et al.
Chemical & Pharmaceutical Bulletin, 29(4), 1131-1131 (1981)
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Protocols

Spectrophotometric Determination of Reinheitszahl (RZ) for Peroxidase (EC 1.11.1.7)

Reinheitszahl (RZ) is the ratio of absorbance due to hemin (A403, Soret region) to absorbance due to protein (A275).

Enzymatic Assay of Peroxidase (EC 1.11.1.7) 2,2'-Azino-bis(3-Ethylbenzthiazoline-6-Sulfonic Acid) as a Substrate

To standardize a procedure for the assay of Peroxidase using 2,2'-Azino-bis(3-Ethylbenzthiazoline-6-Sulfonic Acid) as a substrate.

Enzymatic Assay of Peroxidase (EC 1.11.1.7)

This procedure is for the determination of Peroxidase enzymatic activity using Pyrogallol as the substrate.

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