Gelsolin elisa kit instruction manual
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September 29 06:14:48, 2025
**Gelsolin ELISA Kit Instruction Manual**
**Kit Specifications:**
This Gelsolin ELISA Kit is available in 48-well or 96-well configurations. The kit includes:
- Standard Dilution: 1.5 ml × 1 bottle
- Enzyme Standard Reagent: 3 ml × 1 bottle (for 48-well) or 6 ml × 1 bottle (for 96-well)
**Usage Purpose:**
This reagent is intended solely for research purposes. The standard curve is plotted with the concentration on the x-axis and OD value on the y-axis. Based on the sample's OD value, the corresponding concentration can be determined from the standard curve. Alternatively, a linear regression equation can be calculated using the standard concentrations and their respective OD values. The sample’s OD value is then substituted into the equation to determine its actual concentration, which must be multiplied by the dilution factor.
**Kit Composition:**
- Sealing Film: 2 pieces (for 48-well) / 2 pieces (for 96-well)
- Standard: 0.5 ml × 1 bottle, 2700 ng/L
- Enzyme Standard: 1×48 / 1×96
- Sample Diluent: 3 ml × 1 bottle (for 48-well) or 6 ml × 1 bottle (for 96-well)
- Developer A: 3 ml × 1 bottle (for 48-well) or 6 ml × 1 bottle (for 96-well)
- Chromogen B: 3 ml × 1 bottle (for 48-well) or 6 ml × 1 bottle (for 96-well)
- Wash Solution: 3 ml × 1 bottle (for 48-well) or 6 ml × 1 bottle (for 96-well)
- Concentrated Wash Solution: 20 ml × 20 times (for 48-well) or 20 ml × 30 times (for 96-well)
**Storage Conditions & Expiration Date:**
- Storage: 2–8°C
- Shelf Life: 6 months
**Principle of the Assay:**
The Gelsolin ELISA Kit uses the sandwich method to detect human Gelsolin levels in samples. A microplate is coated with purified anti-Gelsolin antibody. After adding the sample and HRP-labeled Gelsolin antibody, an immune complex is formed. After washing, TMB substrate is added, leading to a color change that is proportional to the Gelsolin concentration. The reaction is stopped, and the absorbance at 450 nm is measured. The sample concentration is determined from the standard curve.
**Sample Preparation & Requirements:**
- **Serum:** Allow blood to clot at room temperature for 10–20 minutes, then centrifuge at 2000–3000 rpm for 20 minutes.
- **Plasma:** Use EDTA or sodium citrate as anticoagulant, mix for 10–20 minutes, then centrifuge.
- **Urine:** Centrifuge at 2000–3000 rpm for 20 minutes.
- **Cell Culture Supernatant:** Centrifuge after collection. For intracellular components, lyse cells by repeated freezing and thawing.
- **Tissue Specimen:** Homogenize in PBS, centrifuge, and collect supernatant.
- **General Notes:** Process samples immediately after collection. If not tested right away, store at -20°C, avoiding repeated freeze-thaw cycles. Avoid using samples containing NaN3.
**Operation Steps:**
1. **Standard Dilution:** Prepare serial dilutions of the standard.
2. **Loading:** Add 40 μl of sample diluent and 10 μl of sample to each well.
3. **Incubation:** Seal the plate and incubate at 37°C for 30 minutes.
4. **Washing:** Rinse wells with diluted wash solution 5 times.
5. **Enzyme Addition:** Add 50 μl of enzyme-labeled reagent to each well.
6. **Second Incubation:** Repeat incubation step.
7. **Color Development:** Add 50 μl of chromogen A and B, incubate at 37°C for 15 minutes.
8. **Stop Reaction:** Add 50 μl of stop solution to each well.
9. **Measurement:** Read OD450 within 15 minutes using a microplate reader.
**Notes:**
- Equilibrate the kit at room temperature before use.
- Avoid cross-contamination by using a new sealing film for each experiment.
- Ensure accurate pipetting and maintain consistent timing during loading.
- Always perform a standard curve and consider diluting high-concentration samples if needed.
- Handle all waste as biohazardous material.
- Do not mix reagents from different batches.
- Follow the manual strictly; results should be based on instrument readings.
**Performance:**
- Correlation coefficient (R) ≥ 0.95
- Intra-assay CV < 9%, Inter-assay CV < 11%
- Detection Range: 0.2 IU/L – 6 IU/L
**Service Commitment:**
We offer free technical support during working hours and sample testing services to ensure optimal experimental results.
**Delivery Period:** From payment to delivery.
This kit is designed for accurate and reliable detection of Gelsolin in various biological samples. Proper handling and following the instructions will ensure consistent and reproducible results.