Description
HRP Detection Reagent (Fluorometric) (BA0173) (BA0173)
The HRP Detection Reagent (Fluorometric) (SKU: BA0173) is a stable, ready-to-use 10-acetyl-3,7-dihydroxyphenoxazine (Ampliflu Red) liquid substrate reagent formulated for the detection of trace amounts of horseradish peroxidase (HRP) activity, for example in ELISA. The reagent reacts with hydrogen peroxide in the presence of a peroxidase to produce a highly fluorescent product measured at 530/590 nm, which may also be visualised under a light source at 530 plus/minus 20 nm. It offers exceptional sensitivity, detecting as little as 0.26 pg of HRP per well in a standard 96-well plate assay, equivalent to 0.008 U/L. A single kit provides sufficient reagent for 500 assays in a 96-well plate format. It is intended for research use only.
| Product Name: | HRP Detection Reagent (Fluorometric) (BA0173) |
| SKU: | BA0173 |
| Detection Method: | Fluorimetric (lambda ex/em = 530/590 nm) |
| Detection Range: | Detects as little as 0.26 pg HRP per well or 0.008 U/L |
| Sample Type: | ELISA and other HRP detection applications |
| Species Reactivity: | All |
| Assay Time: | 30 min |
| Kit Size: | 500 Assays |
| Equipment Required: | Microplate reader |
| Storage: | -20C |
| Shelf Life: | 1 year after receipt |
| Shipping: | Gel Pack |
A stable Ampliflu Red (10-acetyl-3,7-dihydroxyphenoxazine) liquid substrate reagent for the fluorimetric detection of trace horseradish peroxidase activity. In the presence of hydrogen peroxide and a peroxidase, the reagent forms a highly fluorescent product measured at 530/590 nm. The reagent detects as little as 0.26 pg HRP per well (0.008 U/L) and is sufficient for 500 assays in 96-well format.
- Stable liquid substrate reagent for detecting trace horseradish peroxidase activity
- Highly sensitive; detects as little as 0.26 pg HRP per well (0.008 U/L)
- Produces a highly fluorescent product (530/590 nm)
- Fluorescent product may also be visualised under a light source (530 plus/minus 20 nm)
- Sufficient for 500 assays in a 96-well plate format
- Detection of horseradish peroxidase activity in ELISA
- Sensitive fluorimetric quantitation of peroxidase activity
Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
| Step | Procedure |
| 1 | Prior to assay, bring the reagent to room temperature. |
| 2 | Add 1 uL of 3% hydrogen peroxide per 1 mL of HRP Reagent. |
| 3 | For ELISA, use 50 uL of Reagent/H2O2 per well in a 96-well plate, or 20 uL per well in a 384-well plate. |
| 4 | Tap the plate briefly and incubate for 30 minutes at room temperature. If desired, stop the reaction with 10 mM KOH (56 mg KOH in 100 mL H2O). |
| 5 | Read fluorescence at 530/590 nm. |
| Component | Quantity | Storage |
| HRP Reagent | 25 mL | -20C |
| 3% Stabilised H2O2 | 100 uL | -20C |
