Duck BAX ELISA Kit
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September 13 10:53:14, 2025
**Duck BAX ELISA Kit – For the quantitative in vitro determination of Duck Bcl-2-associated X protein (BAX) concentrations in serum, plasma, cerebrospinal fluid, tissue homogenate, and other body fluids. Intended for laboratory research use only. NOT FOR USE IN DIAGNOSTIC PROCEDURES.**
This ELISA kit is designed for the accurate quantification of BAX protein levels in various biological samples. The assay is based on the enzyme-linked immunosorbent assay (ELISA) principle, where a colorimetric reaction occurs in response to the presence of BAX. The intensity of the color developed at 450 nm is directly proportional to the concentration of BAX in the sample.
The kit includes a set of calibration standards that are run alongside the test samples. A standard curve is generated by plotting the optical density (OD) values against known BAX concentrations. This allows for the precise determination of BAX levels in unknown samples by comparing their OD readings to the standard curve.
**Sample Collection and Storage:**
- **Serum:** Use a serum separator tube. Allow samples to clot for 2 hours at room temperature or overnight at 4°C before centrifugation at 2000×g for 20 minutes. Collect and assay immediately or aliquot and store at -20°C. Avoid repeated freeze-thaw cycles.
- **Plasma:** Collect using heparin as an anticoagulant. Centrifuge within 30 minutes at 2000×g for 30 minutes at 2–8°C. Store at -20°C. Avoid repeated freeze-thaw cycles.
- **Cell culture supernatants, tissue homogenates, and other body fluids:** Centrifuge to remove particulates. Assay immediately or aliquot and store at -20°C. Ensure no hemolysis or granulation occurs during processing.
**Materials Required (Not Included):**
- 37°C incubator
- Microplate reader capable of measuring absorbance at 450 nm
- Precision pipettes, disposable tips, and absorbent paper
- Distilled or deionized water
**Reagents Provided (Stored at 2–8°C):**
- Microtiter strip plate (12×8 or 12×4 strips)
- Standard (6 vials, 0.5 ml/vial)
- Sample diluent (6.0 ml or 3.0 ml)
- HRP-Conjugate reagent (10.0 ml or 5.0 ml)
- 20X Wash solution (25 ml or 15 ml)
- Chromogen Solution A (6.0 ml or 3.0 ml)
- Chromogen Solution B (6.0 ml or 3.0 ml)
- Stop solution (6.0 ml or 3.0 ml)
- Closure plate membrane (2 units)
- User manual (1 unit)
- Sealed bags (1 unit)
**Standard Concentrations:** 100, 50, 25, 12.5, 6.25, 3.12 ng/mL. If sample values exceed the highest standard, dilute with sample diluent and repeat the test.
**Important Notes:**
- Do not mix reagents from different kit lots.
- All reagents should be brought to room temperature (20–25°C) before use.
- Do not thaw samples or reagents using water baths.
- Never use beyond expiration date.
- Only use deionized or distilled water for dilutions.
- Keep microtiter plates in sealed bags until ready for use.
- Store unused strips at 2–8°C with desiccant.
- Use fresh pipette tips for each transfer to avoid contamination.
- Treat all samples as potentially infectious.
- Dispose of samples safely after inactivation (allow liquid to stand for 30 minutes).
- Substrate solutions must be handled carefully due to potential contamination.
**Reagent Preparation:**
- Wash solution (1X): Dilute 1 volume of 20X wash solution with 19 volumes of deionized or distilled water. Store at 2–8°C for up to 1 month.
**Assay Procedure:**
1. Prepare all reagents and add 50 µL of standard or sample to appropriate wells (excluding blank well).
2. Cover with adhesive strip and incubate for 60 minutes at 37°C.
3. Wash microtiter plate 4 times manually or automatically.
4. Add 50 µL of chromogen A and 50 µL of chromogen B to each well. Incubate for 15 minutes at 37°C, protected from light.
5. Add 50 µL of stop solution to each well. Read OD at 450 nm.
**Data Interpretation:**
- Plot average OD values of standards vs. concentration.
- Subtract blank OD from all readings.
- Locate OD value on Y-axis and draw a line to the standard curve. Intersect X-axis to determine BAX concentration.
- Intra-assay and inter-assay CV% <15%. Assay range: 3.12–100 ng/mL. Sensitivity: <1.0 ng/mL.
- No significant cross-reactivity observed.
**Storage:** Store at 2–8°C for frequent use; for long-term storage, keep at -20°C for up to six months.
**Note:** Always follow good laboratory practices and ensure proper disposal of biohazardous materials.
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