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Mouse acidic fibroblast growth factor 1 (aFGF-1) elisa kit instruction manual
**Mouse Acidic Fibroblast Growth Factor 1 (aFGF-1) ELISA Kit – Instructions for Use**
This ELISA kit is designed for the quantitative determination of mouse acidic fibroblast growth factor 1 (aFGF-1) in biological samples such as serum, plasma, urine, cell culture supernatants, and tissue homogenates. The kit uses a sandwich ELISA method with high specificity and sensitivity.
**Kit Specifications:**
- **Configuration:** 48-well or 96-well format
- **Standard Dilution:** 1.5 mL × 1 vial
- **Enzyme Standard Reagent:** 3 mL × 1 vial (48-well), 6 mL × 1 vial (96-well)
- **Storage Conditions:** 2–8°C
- **Shelf Life:** 6 months from date of manufacture
**Principle of the Assay:**
The assay is based on the double-antibody sandwich method. A monoclonal antibody specific to aFGF-1 is pre-coated onto the microplate. After incubation with the sample, aFGF-1 binds to the coated antibody. A HRP-conjugated secondary antibody is then added, forming a complex. TMB substrate is used for color development, and the absorbance is measured at 450 nm. The concentration of aFGF-1 in the sample is determined by comparing the OD value to a standard curve.
**Kit Components:**
- Sealing Film: 2 pieces (48-well), 2 pieces (96-well)
- Standard: 0.5 mL × 1 vial (2700 ng/L)
- Enzyme Standard: 3 mL × 1 vial (48-well), 6 mL × 1 vial (96-well)
- Sample Diluent: 3 mL × 1 vial (48-well), 6 mL × 1 vial (96-well)
- Developer A: 3 mL × 1 vial (48-well), 6 mL × 1 vial (96-well)
- Chromogen B: 3 mL × 1 vial (48-well), 6 mL × 1 vial (96-well)
- Wash Buffer: 20 mL × 20 times (48-well), 20 mL × 30 times (96-well)
**Sample Preparation:**
- **Serum/Plasma:** Centrifuge at 2000–3000 rpm for 20 minutes after clotting or anticoagulant addition.
- **Urine:** Centrifuge at 2000–3000 rpm for 20 minutes.
- **Cell Culture Supernatant:** Centrifuge at 2000–3000 rpm for 20 minutes.
- **Tissue Homogenate:** Homogenize in PBS, centrifuge, and collect supernatant.
- **Storage:** Store samples at -20°C if not tested immediately. Avoid repeated freeze-thaw cycles.
**Operation Steps:**
1. Prepare standards by serial dilution.
2. Add samples and standards to the microplate.
3. Incubate at 37°C for 30 minutes.
4. Wash the plate 5 times with wash buffer.
5. Add enzyme conjugate and incubate again.
6. Add TMB substrate and incubate for 15 minutes.
7. Stop the reaction with stop solution.
8. Measure OD at 450 nm.
9. Plot standard curve and calculate sample concentrations.
**Notes:**
- Allow the kit to equilibrate to room temperature before use.
- Do not mix reagents from different batches.
- Use a new pipette tip for each step to avoid cross-contamination.
- Always run standards in duplicate.
- Keep all components away from light and moisture.
- Handle all waste as biohazardous material.
**Performance:**
- **Sensitivity:** 0.2 IU/L
- **Dynamic Range:** 0.2–6 IU/L
- **Correlation Coefficient (R²):** ≥ 0.95
**Technical Support:**
Our team provides free technical assistance during working hours. If you need help with sample testing or interpretation, feel free to contact us. We are committed to ensuring your experiments are successful and accurate.
**Important:** This kit is for research use only. Not for human diagnostic purposes.