10814407001

Peroxidase (POD)

EIA grade, from horseradish

• Synonym(s): POD
• Enzyme Commission number: 1.11.1.7 (BRENDA, IUBMB)
• UNSPSC Code: 12352204

Properties

• biological source: horseradish
• Quality Level: 100
• Assay: 90% (HPLC)
• form: lyophilized (salt-free)
• specific activity: ~225 U/mg lyophilizate, 25 °C (Guaiacol, H2O2)
• packaging: pkg of 100 mg
• manufacturer/tradename: Roche
• concentration: 10 g/L
• technique(s): activity assay: suitable
• color: red to brown
• optimum pH: 6.0-6.5
• solubility: water: soluble at 68 °F
• suitability: suitable for molecular biology
• UniProt accession no.: P00433
• application(s): detection; life science and biopharma
• foreign activity: ATPase <0.00100%, none detected; AcP <0.00100%; Catalase <0.700%
• shipped in: dry ice
• storage temp.: −20°C (−15°C to −25°C)

Description

General description

Donor: hydrogen-peroxide oxidoreductase
Peroxidase can be coupled to other proteins via its amino groups, as well as its carbohydrate moiety. The reconstituted solution can be used directly for conjugation without prior dialysis.

Application

Peroxidase (POD) has been used in the spermine oxidase enzyme activity assay.

Sequence

Blocking substances
Preparations does not contain substances that block the amino groups, but in contrast to other enzymes POD has only 2 to 3 free amino groups which are not accessible; even by extended dialysis of POD, the number of NH₂-groups/mol does not change.

POD
POD contains 1 molecule of protoheme IX as prosthetic group and 2 molecules of Ca²⁺.
POD exists in > 10 isoenzymic forms (some sources estimate approx. 40 isoenzymes).

Carbohydrate Content
12 to 14.5 mg/100 mg Iyophilizate

Free Amino Groups
Free amino groups: 2 to 3 mol/mol enzyme

Preparation Note

Activator: Ammonia, pyridine, imidazole, all at pH >7; detergents
Working concentration: A suggested working ratio, for immediate use in coupling procedures, is 2 mg POD to 4 mg lgG (molar ratio = 2:1, POD:IgG).
Specific activity: approximately 225U/mg lyophilizate (+25°C, guaiacol, H₂O₂) approximately 1,000 U/mg lyophilizate (+25°C, ABTS, H₂O₂, pH 5.0) approximately 3,500 U/mg lyophilizate (+25°C, tetramethylbenzidine, H₂O₂)
Amino groups: 2 - 3moles/mole enzyme

Carbohydrates: 12.0 - 14.5% (w/w)
Isoenzymes: ≥90% isoenzyme C (HPLC)
Purity: single peak (HPLC)
Purity number: 3.0 - 3.5 (A₄₀₃/A₂₇₅)

Storage and Stability

Keep container tightly closed in a dry and well-ventilated place.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Safety Information

• Pictograms: GHS08
• Signal Word: Danger
• Hazard Statements: H317 - H334
• Precautionary Statements: P261 - P280 - P284 - P304 + P340 - P333 + P313 - P342 + P311
• Hazard Classifications: Resp. Sens. 1 - Skin Sens. 1
• Storage Class Code: 11 - Combustible Solids
• WGK: WGK 1
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable