| Product |
Human Eosinophil Peroxidase |
| Product No. |
700-03-002 |
| Origin |
Prepared from human blood that has been shown by certified tests to be negative for HbsAg, HCV and HIV antibodies. |
| CAS Number |
9003-99-0 |
| EC Number |
1.11.1.7 |
| Batch No. |
230410 |
| MW |
70 500 Da |
| 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-1, and the activity should be determined prior to use. |
|
| Test/Analysis |
Specifications |
Results |
| Appearance |
|
solid |
| SDS purity |
|
≥ 95 |
| Reinheitszahl |
A413nm / A280nm |
≥ 1.0 (see UV/vis Spectrum) |
| Solubility |
Calculated with a molar extinction coefficient of 110.000 M-1cm-1 at 413 nm |
Reconstitution of 100 µg with 1000 µL distilled water gives approximately a 1.4 µM solution |
|
| MCD Assay |
100 mM Phosphate buffer pH 7
10 mM NaBr
100 µM MCD (Monochlorodimedon)
100 µM HOOH
10 nM EPO
One unit of halogenation activity is defined as the amount of enzyme that halogenates 1 µmol of MCD min-1 at pH 7.0 and 25 °C. |
results of the linear regression
model: y=a0+a1*x
a1/1 = 1.761
a1/2 = 1.534
a1/3 = 1.692
= 1.662 min-1 = 83.5 µM MCD min-1
117.7 Units mg-1
|
| Guaiacol Assay |
100 mM Phosphate buffer pH 7
100 µM guaiacol
100 µM HOOH
50 nM EPO
One unit of peroxidase activity is defined as the amount of enzyme that oxidises 1 µmol of guaiacol min-1 at pH 7.0 and 25 °C. |
results of the linear regression
Model: y=a0+a1*x
a1/1 = 1.273
a1/2 = 1.362
a1/3 = 1.226
= 1.29 min-1 = 48.5 µM guaiacol min-1
13.75 Units mg-1 |
EPO stock solution was adjusted photometrically and a minimum of 11.7 Units
(= 117 Units per mg) were filled in each vial prepared for lyophilisation.
Solid lyophilisate per vial contains a minimum 100 µg active EPO + marginally amounts of salts and impurities (< 50 µg).
|