Description
Indican Assay Kit (BA0036) (BA0036)
The Indican Assay Kit (SKU: BA0036) provides a quantitative colorimetric method for determining urinary indican (indoxyl sulfate). Indican is a putrefaction product formed by bacterial deconjugation of dietary tryptophan to indole in the small intestine, and urinary indican testing is a useful screening tool for dysbiosis, small intestinal bacterial overgrowth (SIBO), constipation, malabsorption and intestinal mucosal permeability. Whereas the traditional Obermayer test gives only qualitative results, this assay is based on an improved Curzon and Walsh method in which indican reacts with a chromogen. The colour intensity of the product at 480 nm is directly proportional to the indican concentration in the sample, giving a linear range of 0.2 to 20 mg/dL. The homogeneous mix-incubate-measure format takes only ten minutes and is readily automated for processing thousands of samples per day. Both 96-well plate and cuvette protocols are provided.
| Product Name: | Indican Assay Kit (BA0036) |
| SKU: | BA0036 |
| Detection Method: | Colorimetric; improved Curzon and Walsh method with a chromogen measured at 480 nm |
| Detection Range: | 0.2 - 20 mg/dL (8-800 µmoles/L) |
| Sample Type: | Urine |
| Species Reactivity: | All |
| Assay Time: | 10 minutes |
| Kit Size: | 1,000 Assays |
| Equipment Required: | Microplate reader |
| Storage: | Store Reagent A at room temperature and other components at -20°C. |
| Shelf Life: | 12 months after receipt. |
| Shipping: | Room Temperature |
A quantitative colorimetric assay for the determination of urinary indican (indoxyl sulfate). Based on an improved Curzon and Walsh method, indican reacts with a chromogen and the colour intensity of the product at 480 nm is directly proportional to the indican concentration. Linear detection range is 0.2 to 20 mg/dL (8-800 µmoles/L).
- Fast and sensitive: linear detection range 0.2 - 20 mg/dL (8-800 µmoles/L).
- Fast and high-throughput: homogeneous mix-incubate-measure format completed in 10 min; readily automated for thousands of samples per day.
- Quantitative direct determination of indican in urine 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 clear flat-bottom 96-well plates and bring all reagents to room temperature; vortex Reagent B briefly. |
| 2 | Transfer 50 µL urine samples into separate wells of the plate. |
| 3 | Add 140 µL Reagent A to each well, tap to mix and measure OD480nm (OD0). |
| 4 | Add 10 µL Reagent B to each well, tap to mix, incubate 5 min and read OD480nm (ODS). |
| 5 | Add 10 µL provided standard to each well, tap to mix, incubate 5 min and read OD480nm (ODSTD). |
96-well plate assay: [Indican] = [(ODS - OD0) / (ODSTD - ODS)] x 5 x n (mg/dL), where OD0, ODS and ODSTD are the OD480nm values of the sample blank, the endogenous indican and the total indican (endogenous plus 5 mg/dL spiked), respectively, and n is the dilution factor. Conversion: 1 mg/dL indican (indoxyl sulfate potassium salt) = 39.8 µmoles/L, or 10 ppm.
| Component | Quantity | Storage |
| Reagent A | 200 mL | Room temperature |
| Reagent B | 15 mL | -20°C |
| Standard (30 mg/dL) | 15 mL | -20°C |
