Description
Tryptophan Assay Kit (Fluorometric) (BA0153) (BA0153)
The Tryptophan Assay Kit (SKU: BA0153) provides a fast and sensitive fluorometric method for measuring tryptophan concentration. Tryptophan is one of the eight essential amino acids that the body cannot synthesise and must obtain through diet; it is the biochemical precursor to the neurotransmitter serotonin, which regulates appetite, sleep and mood, and to melatonin, which regulates the sleep cycle. This assay uses a coupled enzymatic reaction and the addition of a single working reagent to determine the tryptophan concentration of a sample. The fluorescence intensity at an excitation/emission of 530/585 nm is directly proportional to the tryptophan concentration in the sample. The homogeneous mix-incubate-measure format is readily automated for high-throughput use.
| Product Name: | Tryptophan Assay Kit (Fluorometric) (BA0153) |
| SKU: | BA0153 |
| Detection Method: | Fluorometric |
| Detection Range: | 10 - 400 uM tryptophan |
| Sample Type: | Serum |
| Species Reactivity: | All |
| Assay Time: | 30 minutes |
| Kit Size: | 100 Assays |
| Equipment Required: | Microplate reader |
| Storage: | Store all kit components at -20 C. |
| Shelf Life: | 6 months after receipt |
| Shipping: | Gel Pack |
A coupled enzymatic assay for the quantitative fluorometric determination of tryptophan. A single working reagent generates a fluorescent product measured at 530/585 nm, with intensity proportional to tryptophan concentration. Samples are measured using an internal standard method in three parallel reactions.
- Fast and sensitive, with a linear detection range of 10 - 400 uM tryptophan
- Convenient homogeneous mix-incubate-measure format
- Internal standard method for accurate quantitation in complex matrices
- Readily automated on HTS liquid-handling systems
- Tryptophan determination in serum
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 | Samples require an internal standard and three separate reactions: sample plus standard, sample alone and sample blank. |
| 2 | Prepare 500 uL of 100 uM tryptophan standard (10 uL of 5 mM Standard in 490 uL water). For the sample-plus-standard well add 5 uL 100 uM tryptophan and 10 uL sample; for the sample and sample blank wells add 5 uL water and 10 uL sample. |
| 3 | Prepare Working Reagent per reaction by mixing 105 uL Enzyme Mix, 1 uL Dye Reagent and 1 uL TRP Enzyme. For sample blanks prepare a blank Working Reagent without the TRP Enzyme. |
| 4 | Add 100 uL Working Reagent to each sample-plus-standard and sample-alone well, and 100 uL blank Working Reagent to each sample blank well. Tap to mix. |
| 5 | Incubate for 30 min at room temperature protected from light using a black flat-bottom 96-well plate. |
| 6 | Read fluorescence at an excitation/emission of 530/585 nm. |
Calculate [Tryptophan] = ((F_SAMPLE - F_BLANK) / (F_STANDARD - F_SAMPLE)) x 50 x n (uM), where F values are the fluorescence readings of the sample, sample blank and sample-plus-standard, and n is the dilution factor. If the concentration exceeds 250 uM, dilute the sample in water and repeat. One uM tryptophan equals 0.204 mg/L or 0.204 ppm.
| Component | Quantity | Storage |
| Enzyme Mix | 12 mL | -20 C |
| TRP Enzyme | 120 uL | -20 C |
| Dye Reagent | 120 uL | -20 C |
| Tryptophan Standard (5 mM) | 100 uL | -20 C |
