Horseradish peroxidase (HRP) is isolated from horseradish roots (Amoracia rusticana) and belongs to the ferroprotoporphyrin group of peroxidases. HRP is a single chain polypeptide containing four disulfide bridges. It is a glycoprotein containing 18% carbohydrate. The carbohydrate composition consistsof galactose, arabinose, xylose, fucose, mannose, mannosamine, and galactosamine, depending upon the specific isozyme.2 Its molecular weight (~44kD) includes the polypeptide chain (33,890 Daltons), hemin plus Ca2+ (approx. 700 Daltons), and carbo-hydrate (9400 Daltons).3 At least seven isozymes of HRP exist.2 The isoelectric point for horseradish Peroxidase isozymes ranges from 3-9. HRP readily combines with hydrogen peroxide (H2O2) and the resultant [HRP-H2O2] complex can oxidize a wide variety of chromogenic hydrogen donors. It can also utilize chemiluminescent substrates such as luminol and isoluminol and fluorogenic substrates such as tyramine, homovanillic acid, 4-hydroxyphenyl acetic acid.
The following compounds are inhibitors of horseradish peroxidase: sodium azide, cyanide, L-cystine, dichromate, ethylenethiourea, hydroxylamine, sulfide, vanadate, paminobenzoic acid, Cd+2, Co+2, Cu+2, Fe+3, Mn+2, Ni+2, Pb+2. 4 The pH optimum of HRP is in the range of 6.0 to 6.5; activity at 7.5 is 84% of the maximum. The enzyme is most stable in the pH range of 5.0 to 9.0.5 Horseradish peroxidase is widely used as a label for immunoglobulins in many different immunochemistry leads to lower non-specific binding.6 Protocols describing the glutaraldehyde and periodate conjugation methodologies can be reviewed in Harlow, E. et al.
Source:HRP, from horseradish roots (Amoracia rusticana)
RZ (Reinheitszahl):Absorbance ratio of A403/A275. RZ=3.0.
Purpurogallin Activity (units per mg solid):~250-330
Purpurogallin Unit Definition:One unit will form 1.0mg purpurogallin from pyrogallol in 20 seconds at pH 6.0 at 20 °C. This unit is equivalent to approx. 18uM units/minute at 25 °C.
ABTS Unit Definition:One unit will oxidize 1umole of 2, 2’-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid)/minute at 25°C, pH 5.0.
ABTS Activity (units per mg solid):~1000
Storage and Stability:Lyophilized powder may be stored at -20°C. Stable for 12 months after receipt at -20°C. Reconstitute with sterile buffer or ddH2O. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Reconstituted product is stable for 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
仕様
Size：1mg
Source Antigen：Horseradish roots
Grade：Purified
Purity：Purified
Form：Supplied as a lyophilized powder. The choice of solvent will depend on the intended application. The powdered enzymes are soluble in H2O or 0.1M phosphate buffer, pH 6 (10mg/ml).
