Gelsolin elisa kit instruction manual
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September 29 06:11:02, 2025
**Gelsolin ELISA Kit Instruction Manual**
**Kit Specifications:**
The Gelsolin ELISA Kit is available in 48-well or 96-well configurations. The standard dilution is 1.5 ml × 1 vial. The enzyme standard reagent is provided as 3 ml × 1 vial (for 48-well) or 6 ml × 1 vial (for 96-well). This reagent is for research use only. To determine the concentration of Gelsolin, plot the standard concentrations on the x-axis and the corresponding OD values on the y-axis. Draw a standard curve and calculate the sample concentration based on its OD value. Alternatively, use the linear regression equation derived from the standard curve to calculate the sample concentration. Multiply by the dilution factor to obtain the actual sample concentration.
**Kit Components:**
- Sealing film: 2 pieces (48) / 2 pieces (96)
- Standard: 2700 ng/L, 0.5 ml × 1 bottle
- Enzyme standard: 1×48 / 1×96
- Sample diluent: 3 ml × 1 bottle (48) / 6 ml × 1 bottle (96)
- Developer A: 3 ml × 1 bottle (48) / 6 ml × 1 bottle (96)
- Chromogen B: 3 ml × 1 bottle (48) / 6 ml × 1 bottle (96)
- Wash buffer: 3 ml × 1 bottle (48) / 6 ml × 1 bottle (96)
- Concentrated wash solution: 20 ml × 20 times (48) / 20 ml × 30 times (96)
**Storage Conditions & Expiration:**
- Store at 2–8°C
- Shelf life: 6 months
**Principle of the Assay:**
This kit uses a sandwich ELISA method to detect human Gelsolin in serum, plasma, urine, cell culture supernatants, and tissue homogenates. The microplate is pre-coated with anti-Gelsolin antibodies. After adding the sample and HRP-labeled Gelsolin antibody, an immune complex forms. Unbound components are washed away, and TMB substrate is added. The color develops and turns yellow upon acid termination. The intensity of the color is proportional to the Gelsolin concentration. The absorbance is measured at 450 nm, and the concentration is calculated using a standard curve.
**Purpose:**
To quantify Gelsolin levels in biological samples such as serum, plasma, urine, and tissue homogenates.
**Sample Preparation Guidelines:**
1. **Serum:** Allow blood to clot at room temperature for 10–20 minutes, then centrifuge at 2000–3000 rpm for 20 minutes. Collect the supernatant.
2. **Plasma:** Use EDTA or sodium citrate as anticoagulant. Centrifuge after mixing for 10–20 minutes.
3. **Urine:** Centrifuge at 2000–3000 rpm for 20 minutes.
4. **Cell Culture Supernatant:** Centrifuge at 2000–3000 rpm for 20 minutes. For intracellular components, lyse cells via freeze-thaw cycles before centrifuging.
5. **Tissue:** Homogenize in PBS (pH 7.4), centrifuge, and collect the supernatant.
6. **General Note:** Process samples as soon as possible. If not tested immediately, store at -20°C. Avoid repeated freezing and thawing. Do not use samples containing NaN3.
**Operation Steps:**
1. **Standard Dilution:** Prepare a 5-point standard curve (1800, 1200, 600, 300, 150 ng/L).
2. **Sample Loading:** Add 40 μl of sample diluent and 10 μl of sample into each well.
3. **Incubation:** Seal the plate and incubate at 37°C for 30 minutes.
4. **Washing:** Use diluted washing solution (1:30) and wash 5 times.
5. **Enzyme Addition:** Add 50 μl of enzyme-labeled reagent to each well.
6. **Incubation:** Repeat step 3.
7. **Color Development:** Add 50 μl of TMB A and B, incubate at 37°C for 15 minutes.
8. **Stop Reaction:** Add 50 μl of stop solution.
9. **Measurement:** Read OD at 450 nm within 15 minutes.
**Notes:**
- Equilibrate the kit at room temperature for 15–30 minutes before use.
- Avoid cross-contamination; use a new sealing film for each test.
- Handle all materials as biohazardous waste.
- Do not mix reagents from different batches.
- Always perform a standard curve and double wells for accuracy.
- Ensure accurate pipetting and avoid light exposure during TMB development.
**Performance:**
- Correlation coefficient (R²) ≥ 0.95
- Intra-batch CV < 9%, Inter-batch CV < 11%
- Detection range: 0.2 IU/L – 6 IU/L
**Service Commitment:**
We offer free technical support during working hours. Sample testing services are available upon request.
**Delivery:** From payment to delivery.
**Note:** This manual is intended for reference. In case of discrepancies, the English version shall prevail.