Description
Total Protein Assay Kit (BA0174) (BA0174)
The Total Protein Assay Kit (SKU: BA0174) offers a simple, direct and automation-ready method for the fluorimetric determination of protein or peptide concentration in a wide range of biological samples. Based on an improved o-phthalaldehyde method, the reagent reacts with the primary amines of proteins and peptides to form a blue fluorescent product, allowing the detection of nanogram quantities of protein. The fluorescence intensity, measured at 360/450 nm, is proportional to the protein concentration in the sample. The assay is completed within a few minutes, offers a linear detection range of 1.3-1000 ug/mL BSA and is compatible with various lysis buffers, making it ideal for determining protein concentrations in cell or tissue lysates. This homogeneous mix-incubate-measure format can be readily automated for high-throughput processing. It is intended for research use only.
| Product Name: | Total Protein Assay Kit (BA0174) |
| SKU: | BA0174 |
| Detection Method: | Fluorimetric (lambda ex/em = 360/450 nm) |
| Detection Range: | 1.3 - 1000 ug/mL BSA |
| Sample Type: | Cell and tissue lysates and other biological samples |
| Species Reactivity: | All |
| Assay Time: | 5 min |
| Kit Size: | 200 Assays |
| Equipment Required: | Microplate reader |
| Storage: | -20C |
| Shelf Life: | 6 months after receipt |
| Shipping: | Room Temperature |
A fast, sensitive fluorimetric assay for the quantitative determination of protein or peptide concentration based on an improved o-phthalaldehyde method. The reagent reacts with primary amines to form a blue fluorescent product measured at 360/450 nm, proportional to protein concentration, with a linear range of 1.3-1000 ug/mL BSA. The homogeneous assay works directly with various lysis buffers and is completed within a few minutes.
- Fast and sensitive; assay completed within a few minutes
- Linear detection range of 1.3 - 1000 ug/mL BSA
- Compatible with various lysis buffers, ideal for cell and tissue lysates
- Homogeneous mix-incubate-measure format with no wash or reagent-transfer steps
- Readily automated on high-throughput liquid-handling systems
- Quantitative determination of protein in various biological samples
- Measurement of protein concentration in cell and tissue lysates
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 and use black flat-bottom 96-well plates. Prepare a 1000 ug/mL Standard Premix by mixing 10 uL of the 20 mg/mL Standard with 190 uL H2O, then dilute as shown in the table. |
| 2 | Transfer 10 uL of each standard and 10 uL of each sample into separate wells of the plate (use the same buffer concentration instead of H2O for lysate samples). |
| 3 | Add 90 uL Reagent to all wells and immediately tap the plate to mix. |
| 4 | Incubate for 5 minutes at room temperature. |
| 5 | Measure fluorescence intensity at 360/450 nm on a plate reader, reading all samples at the same time interval after mixing. |
Plot the protein standard curve and determine its slope. [Protein] = (F_SAMPLE - F_BLANK) / Slope (ug/mL), where F_SAMPLE and F_BLANK are the fluorescence values of the sample and the blank (#4 H2O), respectively.
| Component | Quantity | Storage |
| Reagent | 20 mL | -20C |
| Standard (20 mg/mL BSA) | 100 uL | -20C |
