Description
Total Protein (Urine/CSF) Assay Kit (BA0179) (BA0179)
The Total Protein (Urine/CSF) Assay Kit (SKU: BA0179) provides a simple, reliable and rapid colorimetric method for the quantitative determination of total protein, particularly for assessing renal and blood-brain barrier functions. The assay is based on an improved pyrogallol red-molybdate protein dye-binding method, in which the colour intensity at 600 nm is directly proportional to the total protein concentration in the sample. This single reagent can be used to determine total protein in urine (proteinuria) and cerebrospinal fluid (blood-brain barrier). The assay takes only 10 minutes, offers a linear detection range of 5-200 mg/dL (1-20 mg/dL for urine samples using an internal standard) and shows minimal interference (< 2%) from substances present in urine. The homogeneous mix-incubate-measure format can be readily automated for high-throughput processing. It is intended for research use only.
| Product Name: | Total Protein (Urine/CSF) Assay Kit (BA0179) |
| SKU: | BA0179 |
| Detection Method: | Colorimetric (OD600 nm) |
| Detection Range: | 5 - 200 mg/dL protein; 1 - 20 mg/dL protein for urine samples |
| Sample Type: | Urine and cerebrospinal fluid |
| Species Reactivity: | All |
| Assay Time: | 10 min |
| Kit Size: | 100 Assays |
| Equipment Required: | Microplate reader |
| Storage: | 2-8C |
| Shelf Life: | 12 months after receipt |
| Shipping: | Room Temperature |
A rapid colorimetric assay for the quantitative determination of total protein based on an improved pyrogallol red-molybdate dye-binding method. The colour intensity measured at 600 nm is directly proportional to the total protein concentration, with a linear range of 5-200 mg/dL (1-20 mg/dL for urine samples using an internal standard). The single reagent is suited to determining protein in urine and cerebrospinal fluid, with minimal interference (< 2%) from urine substances.
- Fast and sensitive; assay takes 10 minutes with a linear range of 5 - 200 mg/dL protein (1 - 20 mg/dL for urine)
- Robust and reliable; substances in urine have minimal interference (< 2%)
- Convenient homogeneous mix-incubate-measure format
- Readily automated on high-throughput liquid-handling systems
- Determination of total protein in urine (proteinuria)
- Determination of total protein in cerebrospinal fluid (blood-brain barrier assessment)
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 | Bring all reagents to room temperature. Standard procedure: pipette 5 uL dH2O, 5 uL provided Protein Standard and 5 uL samples into separate wells of a clear flat-bottom 96-well plate. |
| 2 | Add 200 uL Reagent to all assay wells and tap the plate to mix. |
| 3 | Incubate for 10 minutes and measure OD600nm on a plate reader. |
| 4 | Urine samples: dilute the 100 mg/dL Standard to 40 mg/dL, run samples in duplicate, and transfer 20 uL of each sample into two Sample wells and two Internal Standard wells; add 5 uL dH2O to Sample wells and 5 uL of the 40 mg/dL Standard to Internal Standard wells, with 25 uL dH2O as the Blank in duplicate. |
| 5 | For urine, add 200 uL Reagent to each protein-determination well, incubate 10 minutes at room temperature and read OD at 600 nm. |
Standard samples: [Protein] = [(ODSTANDARD - ODH2O) / (ODSAMPLE - ODH2O)] x 100 x n (mg/dL). Internal-standard samples (e.g. urine): [Protein] = [(ODSTANDARD - ODSAMPLE) / (ODSAMPLE - ODBLANK)] x 10 x n (mg/dL). If the calculated protein concentration exceeds 200 mg/dL, dilute the sample in water and repeat the assay, multiplying the result by the dilution factor n.
| Component | Quantity | Storage |
| Reagent | 20 mL | 2-8C |
| Standard (100 mg/dL) | 1 mL | 2-8C |
