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product specification of

 

human eosinophil peroxidase

 

 

General Product Information


Product name: Eosinophil Peroxidase from human eosinophils
Description:

Haem protein that catalyses oxidations by H2O2, including EPO-bromide-mediated killing of helmintic parasites, metazoan pathogens and cancer cells, and the enzyme is associated with the majority of tissue damage in asthma.

Form:

Brown solid, lyophilized from 5 mM phosphate buffer, pH 7.0.

Solubility:

Reconstitution of 100 µg with 800 µl distilled water gives approximately 1.7 µM EPO calculated with a molar extinction coefficient at 413 nm of 110 000 M-1cm-1.

Storage:

Stored below 0°C this lyophilized product is stable for years.
Storage of the reconstituted form at 4°C for 2 weeks has no effect on enzyme activity. If solutions must be stored for extended periods of time, protein concentration should be kept above 1 mg/ml, and the activity should be determined prior to use.

Source:

Prepared from human blood that has been shown by certified tests to be negative for HBsAg and HIV antibodies.

FOR RESEARCH ONLY!
NOT FOR HUMAN OR DRUG USE!

EC Number:

1.11.1.7

Molecular weight:

~ 70 500 Da    ( see MALDI-TOF Spectrum )

Purity:

> 99%

Reinheitszahl:

A413 / A280 ³ 0.90  ( see UV / VIS Spectrum )
Specific activity:

270
1520

–
–

300
1590

units per mg (assay 1,  Guaiacol)
units per mg (assay 2,  MCD)

for further information about the assays have a look at detailed unit definition


Detailed Unit Definition



 

Assay 1
(Guaiacol)		 Assay 2
(MCD)

 

Assay 1 (Guaiacol):


One unit will produce an increase in absorbance at 470 nm of 1.0 per minute at pH 7.0 and 25°C, calculated from the initial linear rate of reaction using guaiacol as substrate for the peroxidatic activity.

 

Total reaction volume:

1 ml

 

Final assay concentrations:

100 mM phosphate buffer, pH 7.0
100 µM guaiacol
100 µM hydrogen peroxide
 50  nM EPO

 

References:

Desser, R.K., Himmelhoch, S.R., Evans, W.H., Januska, M., Mage, M., and Shelton, E. (1972) Arch. Biochem. Biophys. 148, 452.

 

 

 

 

 

 

 

Assay 1
(Guaiacol)		 Assay 2
(MCD)

 

Assay 2 (MCD):


One unit will produce a decrease in absorbance at 290 nm of 1.0 per minute at pH 7.0 and 25°C, calculated from the initial linear reaction rate using monochlorodimedon (MCD) and bromide as substrate for the brominating activity.

 

Total reaction volume:

1 ml

 

Final assay concentrations:

100 mM phosphate buffer, pH 7.0
100 µM MCD
10   mM NaBr
100 µM hydrogen peroxide
10   nM EPO

 

References:

Hager, L.P., Morris, D.R., Brown, F.S., and Eberwein, H. (1966) J. Biol. Chem. 241, 1769.

Kettle, A.J., and Winterbourn, C.C. (1988) Biochim. Biophys. Acta 957, 185.

 

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