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Total Bile Acid (TBA) Colorimetric Assay Kit
SKU: E-BC-K181-M-500
Total Bile Acid (TBA) Colorimetric Assay Kit
| SKU # | E-BC-K181-M |
| Detection Instrument | Microplate reader (400-410 nm) |
| Detection method | Colorimetric method |
Product Details
Properties
| Synonyms | TBA |
| Sample type | serum, animal tissue |
| Sensitivity | 2.05 μmol/L |
| Detection range | 2.05-120 μmol/L |
| Detection method | Colorimetric method |
| Assay type | Quantitative |
| Assay time | 25 min |
| Precision | Average inter-assay CV: 3.700%Average intra-assay CV: 3.400% |
| Other instruments required | Micropipettor, Centrifuge, Incubator, Vortex mixer |
| Other reagents required | Normal saline (0.9% NaCl) |
| Storage | 2-8℃ |
| Valid period | 12 months |
Images
BB Liu et al investigate the function of SHCZ in ameliorating liver injury. Total Bile Acid (TBA) in rat liver was determined using TBA colorimetric assay kit (E-BC-K181-M).
The concentration of TBA was significantly changed in each group. (*P<0.05, **P<0.01 vs Normal, ##P<0.01 vs Model, △△P<0.01, △△△P<0.001 vs High-dose)
Detection Principle
With S-NAD+ as hydrogen receptor, 3α-hydroxy steroid dehydrogenase catalyzed the dehydrogenation of bile acids to produce 3-ketone steroids, transforming S-NAD+ into S-NADH. Meanwhile, NADH was used as hydrogen donor. 3α-hydroxy steroid dehydrogenase catalyzed the production of bile acids from 3-ketone steroids. Through the enzyme cycle reaction, S-NADH is continuously generated, which has the maximum absorption peak at 405 nm. Measure the OD value at 405 nm and the changes of absorbance is proportional to the concentration of bile acid.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Chromogenic Agent A | 12 mL × 1 vial | 24 mL × 1 vial | 2-8°C, 12 months, shading light |
| Reagent 2 | Chromogenic Agent B | 3 mL × 1 vial | 6 mL × 1 vial | 2-8°C, 12 months, shading light |
| Reagent 3 | 0.2 mmol/L Standard | 2 mL × 1 vial | 2 mL × 1 vial | 2-8°C, 12 months |
| Microplate | 96 wells | No requirement | ||
| Plate Sealer | 2 pieces | |||
Note: The reagents must be stored strictly according to the preservation conditions in the above table. The reagents in different kits cannot be mixed with each other. For a small volume of reagents, please centrifuge before use, so as not to obtain sufficient amount of reagents.
Technical Data:
Parameter:
Intra-assay Precision
Three human serum samples were assayed in replicates of 20 to determine precision within an assay. (CV = Coefficient of Variation)
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (μmol/L) | 5.60 | 13.50 | 34.00 |
| %CV | 3.7 | 3.4 | 3.1 |
Inter-assay Precision
Three human serum samples were assayed 20 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (μmol/L) | 5.60 | 13.50 | 34.00 |
| %CV | 3.1 | 3.8 | 4.2 |
Recovery
Take three samples of high concentration, middle concentration and low concentration to test the samples of each concentration for 6 times parallelly to get the average recovery rate of 97%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc. (μmol/L) | 8.5 | 22 | 33.5 |
| Observed Conc. (μmol/L) | 8.6 | 20.7 | 32.2 |
| Recovery rate (%) | 101 | 94 | 96 |
Sensitivity
The analytical sensitivity of the assay is 1.36 μmol/L. This was determined by adding two standard deviations to the mean O.D. obtained when the zero standard was assayed 20 times, and calculating the corresponding concentration.
Standard Curve
As the OD value of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique or temperature effects), so the standard curve and data are provided as below for reference only.
| Concentration (μmol/L) | 0 | 5 | 10 | 20 | 25 | 30 | 35 | 40 |
| A1 value | 0.348 | 0.384 | 0.421 | 0.487 | 0.530 | 0.578 | 0.607 | 0.644 |
| 0.349 | 0.384 | 0.422 | 0.492 | 0.535 | 0.575 | 0.604 | 0.648 | |
| Average A1 | 0.349 | 0.384 | 0.422 | 0.490 | 0.533 | 0.577 | 0.606 | 0.646 |
| A2 value | 0.350 | 0.452 | 0.557 | 0.743 | 0.850 | 0.978 | 1.044 | 1.166 |
| 0.350 | 0.452 | 0.559 | 0.748 | 0.872 | 0.970 | 1.055 | 1.179 | |
| Average A2 | 0.350 | 0.452 | 0.558 | 0.746 | 0.861 | 0.974 | 1.050 | 1.173 |
| ΔA/min = (A2-A1)/5 | 0.000 | 0.014 | 0.027 | 0.051 | 0.066 | 0.080 | 0.089 | 0.105 |
| Absoluted ΔA/min | 0.000 | 0.013 | 0.027 | 0.051 | 0.066 | 0.079 | 0.089 | 0.105 |