Merck, 10814407001, Peroxidase (POD)

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

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