Horseradish Peroxidase
HRP-Juice, single bottle
Horseradish peroxidise enzyme is the most widely used in EIA and EIH. Typical peroxidases are hemoproteins and transfer hydrogen from hydrogen donors to hydrogren peroxide. Horseradish peroxidises often occur as multiple isozymes and are widely distributed in plants.
Three main types of horseradish peroxidises have been identified, (i) the acidic horseradish peroxidise with a very high carbohydrate content, (ii) horseradish peroxidises with a neutral pH with lower sugar content, (iii) very basic horseradish peroxidises (pI>11) of low sugar content.
Introduction
Enzyme conjugates are used in enzyme immunoassays (EIA), enzyme-linked immunosorbent assays (ELISA), blotting techniques, reporter gene assays, in-situ hybridisation cytometry and histometry. Most frequently Horseradish Peroxidase (HRP), Alkaline Phosphatase (AP) and b-Galactosidase enzymes are used because of their high turnover rate, stability, easy of conjugation and relatively low cost.
Chemiluminescent and Bioluminescent reactions are becoming increasing important in ultra sensitive assays for clinically significant substance i.e. hormones, cancer markers, viral antibodies and in the study of biological process. Two notable recent development are the high sensitivity assays for HRP and AP labels. The peroxidise assay is based on enhanced chemiluminescent luminol reactions and the alkaline phosphatase assay utilizes a stabilized chemiluminescent 1,2-dioxetane phosphate substrate which undergoes an enzyme catalyzed decomposition to produce a characteristic chemiluminescence emission.
Useage:
Do not contaminate the HRP enzyme substrate with HRP enzyme or other proteins. HRP enzyme substrate has a wide range to detect HRP enzyme in solution as well as on membrane.The lower enzyme concentration may be a few femto grams and the highest concentration HRP of enzyme may be one nano gram depending on the source of the HRP enzyme. For good results, wash your tube or micro titer plate or membrane with 0.2M phosphate buffer, pH 8.4 to 8.6, and then use the substrate. Best results for chemiluminescence can be obtained from 6 minutes to 30 minutes after contacting substrate with HRP enzyme..
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HRP-Juice, two bottle
Our stabilized ultra-sensitive Luminol based substrates detect horseradish peroxidase at femtogram level and our chemiluminescent systems 3 to 5 more times sensitive than our best competitor. These reagents are stable for 21 days after mixing when stored at 4°C.
Click here for the Standard Protocol HRP-Juice, two bottle
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Additional Informations
Storage: Store the reagent bottles, HRP-A and HRP-B, at 4-8 °C .
Usage: Mix equal volumes of HRP-A and HRP-B in a clean container and equilibrate at room temperature for 30 minutes before use. Do not contaminate the HRP enzyme substrate with HRP enzyme or other proteins.HRP enzyme substrate has a wide range to detect HRP enzyme in solution as well as in ELISA. The lower enzyme concentration may be a few femto grams and the highest concentration HRP of enzyme may be one nano gram depending on the source of the HRP enzyme.
For good results, wash your tube or micro titer plate or membrane with 0.2M phosphate buffer, pH 8.4 to 8.6, and then use the substrate. Best results for chemiluminescence can be obtained from 2 minutes to 20 minutes after contacting substrate with HRP enzyme.