BioReagent BioReagent for sensitive chromatographic and analytical workflows requiring minimal baseline interference.
Store at 2-8°C,Protected from light Ships Wet ice Check lot-specific COA for exact specifications.
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Cited in 0 peer-reviewed publications across chromatography, organic synthesis, and cross-coupling reactions.
TBA is catabolized in the liver, and elevated serum TBA concentrations indicate parenchymal liver injury. Therefore, TBA measurement is highly valuable for monitoring chronic liver diseases. Bile acids are specifically oxidized by 3α-hydroxysteroid dehydrogenase (3α-HSD) and oxidized β-thionicotinamide adenine dinucleotide (Thio-NAD) to produce 3-ketosteroids and reduced β-thionicotinamide adenine dinucleotide (Thio-NADH). The generated 3-ketosteroids are converted back to bile acids and oxidized β-nicotinamide adenine dinucleotide (NAD) in the presence of 3α-hydroxysteroid dehydrogenase and reduced β-thionicotinamide adenine dinucleotide (Thio-NADH). The cyclic amplification reaction described above improves assay sensitivity. The absorbance change of the produced reduced β-thionicotinamide adenine dinucleotide (Thio-NADH) at 405 nm within a unit time is measured to calculate the bile acid concentration.
Note: This kit is for research use only, and shall not be applied for clinical diagnosis or other purposes.
Required Instruments and Reagents (to be supplied by user)
⁕Full-wavelength microplate reader (capable of measuring absorbance at 405 nm)
⁕Mortar (homogenizer), ice box (ice maker), tabletop centrifuge, adjustable pipettes, water bath (drying oven, incubator, metal bath), 96-well plates, centrifuge tubes
⁕Distilled water (deionized water or ultrapure water is acceptable)
Assay Protocol:
It is recommended to select 1–3 samples with significant differences (e.g., different types or groups) for a pilot experiment to get familiar with the operating procedure. Adjust sample concentration according to pilot results to avoid unnecessary waste of samples and reagents!
1. Sample Extraction:
① Liquid samples: Clear liquid can be tested directly; turbid liquid shall be centrifuged and the supernatant collected for detection.
② Tissue samples: Weigh approximately 0.1 g tissue into a mortar, add 1 mL absolute ethanol, and homogenize on ice. Make up the final volume to 1 mL with absolute ethanol if liquid loss occurs. Centrifuge at 12,000 rpm for 10 min at 4 °C or room temperature, and collect the supernatant for testing.
③ Cell samples: Harvest cells into centrifuge tubes and discard supernatant after centrifugation. Add roughly 5 million bacteria or cells into 1 mL absolute ethanol, then lyse cells/bacteria by sonication on ice (power: 200 W, sonicate for 3 s, pause for 10 s, repeat 30 times). Centrifuge at 12,000 rpm, 4 °C for 10 min, transfer supernatant and keep on ice prior to measurement.
Note: If larger sample volume is needed, perform extraction at a ratio of 500~1000 : 1 (10⁴ bacteria/cells : mL extract solution).
2. Detection Procedure
① Preheat the microplate reader for 30 min (or wait for the instrument self-check process), set temperature to 37 °C and wavelength to 405 nm.
② Thaw all reagents to room temperature, then add reagents sequentially into the 96-well plate as follows:
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Notes:
1. If the value of A₂ is greater than 0.8, dilute the sample supernatant with normal saline or distilled water, and substitute the dilution factor D into the calculation formula.
2. If the value of ΔA is less than 0.005, you may increase the sample loading volume V₁ (e.g., increase from 10 μL to 20 μL; add 20 μL distilled water to the blank tube instead of 10 μL; for the standard tube, add 10 μL standard solution plus 10 μL distilled water to reach a total volume of 20 μL consistent with the assay tube and blank tube; all other reagent volumes remain unchanged). Alternatively, increase the weighed mass W of the tissue sample. Recalculate using the adjusted V₁ or W in the formula.
Calculation of Results:
1. Calculation by volume:
Total Bile Acid (TBA) (μmol/L) = (Cstd × V₂) × (ΔAtest − ΔAblank) ÷ (ΔAstd − ΔAblank) ÷ V₁ × D = Cstd × (ΔAtest − ΔAblank) ÷ (ΔAstd − ΔAblank) × D
2. Calculation by protein concentration:
Total Bile Acid (TBA) (nmol/mg prot) = (Cstd × V₂) × (ΔAtest − ΔAblank) ÷ (ΔAstd − ΔAblank) ÷ (V₁ ÷ V × Cpr) × D = Cstd × (ΔAtest − ΔAblank) ÷ (ΔAstd − ΔAblank) ÷ Cpr × D
3. Calculation by sample weight:
Total Bile Acid (TBA) (nmol/g tissue) = (Cstd × V₂) × (ΔAtest − ΔAblank) ÷ (ΔAstd − ΔAblank) ÷ (V₁ ÷ V × W) × D = Cstd × (ΔAtest − ΔAblank) ÷ (ΔAstd − ΔAblank) ÷ W × D
4. Calculation by cell count:
Total Bile Acid (TBA) (nmol/10⁴ cells) = (Cstd × V₂) × (ΔAtest − ΔAblank) ÷ (ΔAstd − ΔAblank) ÷ (V₁ ÷ V × 500) × D = Cstd × (ΔAtest − ΔAblank) ÷ (ΔAstd − ΔAblank) ÷ 500 × D
Cstd — Concentration of standard solution
V₁ — Volume of loaded sample, 0.001 mL
V₂ — Volume of loaded standard solution, 0.001 mL
V — Total volume of extraction solution, 1 mL
500 — Cell count unit: ten thousand cells
W — Weighed mass of tissue sample, g
D — Dilution factor; set as 1 if no dilution is performed
Cpr — Protein concentration of sample, mg/mL
| Item No. | Appearance | Components | 96T | Storage |
| T1522089A | Liquid | Reagent 1 | 20 mL | 2-8℃. Store in the dark. |
| T1522089B | Liquid | Reagent 2 | 5 mL | 2-8℃. Store in the dark. |
| T1522089C | Liquid | Standard Tube | 0.2mL | 2-8℃. Store in the dark. |
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| Lot Number | Certificate Type | Datum | Artikel |
|---|---|---|---|
 ZJ26F0737034 | Certificate of Analysis | Aug 14, 2026 | T1522089 |
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