Description
Ammonia Assay Kit (Fluorometric) (BA0054) (BA0054)
The Ammonia Assay Kit (Fluorometric) (SKU: BA0054) provides a fast, sensitive and automation-ready method for the quantitative fluorimetric determination of ammonia and ammonium concentration in biological and environmental samples. Ammonia, or its ionic form ammonium, is an important source of nitrogen for living systems and is ubiquitous in nature. The assay is based on an improved o-phthalaldehyde method, in which the reagent reacts with ammonia and ammonium to form a fluorescent product. The fluorescence intensity, measured at excitation/emission wavelengths of 360/450nm, is proportional to the ammonia concentration in the sample. The homogeneous mix-incubate-measure format can be readily automated on high-throughput liquid handling systems for processing thousands of samples per day, and the assay is compatible with most detergents, chelators and buffer components.
| Product Name: | Ammonia Assay Kit (Fluorometric) (BA0054) |
| SKU: | BA0054 |
| Detection Method: | Fluorometric |
| Detection Range: | 0.012 - 1 mM ammonia |
| Sample Type: | Biological (e.g. urine) and environmental samples |
| Species Reactivity: | All |
| Assay Time: | 15 minutes |
| Kit Size: | 200 Assays |
| Equipment Required: | Microplate reader |
| Storage: | The kit is shipped at room temperature. Store the kit at -20°C. |
| Shelf Life: | 12 months after receipt |
| Shipping: | Room Temperature |
This kit is based on an improved o-phthalaldehyde method, in which the reagent reacts with ammonia and ammonium to form a fluorescent product. The fluorescence intensity (λex/em = 360/450nm) is proportional to the ammonia concentration in the sample.
- Fast and sensitive: linear detection range 0.012 - 1 mM ammonia
- Convenient and high-throughput: homogeneous mix-incubate-measure assay, readily automated on HTS liquid handling systems for thousands of samples per day
- Ammonia/ammonium determination in biological (e.g. urine) and environmental samples
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 | Use black flat-bottom 96-well plates and bring all reagents to room temperature before assay. |
| 2 | Standards: prepare 200 µL 1 mM Standard Premix by mixing 10 µL 20 mM NH4Cl Standard and 190 µL H2O, then dilute as shown in the table. Transfer 10 µL standards into separate wells and 10 µL of each sample into separate wells. |
| 3 | Prepare Working Reagent (per reaction: 90 µL Assay Buffer, 4 µL Reagent A and 4 µL Reagent B) and add 90 µL to all wells; immediately tap plate to mix. |
| 4 | Incubate 15 minutes in the dark at room temperature and measure fluorescence intensity at 360/450nm on a plate reader. |
Plot the ammonia standard curve and determine its slope. Ammonia concentration = (FSAMPLE − FBLANK) / Slope (mM), where FSAMPLE and FBLANK are the fluorescence intensity values of the Sample and the blank (#4, H2O). If ammonia exceeds 1 mM, dilute the Sample in water, repeat the assay and multiply by the dilution factor.
| Component | Quantity | Storage |
| Assay Buffer | 20 mL | -20°C |
| Standard (NH4Cl) | 400 µL | -20°C |
| Reagent A | 1 mL | -20°C |
| Reagent B | 1 mL | -20°C |
