**Human FGF-9 ELISA Kit – For the quantitative in vitro determination of Human fibroblast growth factor-9 concentrations in serum, plasma, cerebrospinal fluid, tissue homogenate, and other body fluids. Intended for laboratory research use only, not for diagnostic or therapeutic procedures.**
This ELISA kit is designed for the accurate and reliable measurement of FGF-9 levels in various biological samples. The assay is based on the enzyme-linked immunosorbent assay (ELISA) principle, utilizing a sandwich format with specific antibodies to capture and detect FGF-9.
The Stop Solution changes the color from blue to yellow, and the optical density (OD) is measured at 450 nm using a microplate reader. A standard curve is generated using a set of calibration standards, allowing for precise quantification of FGF-9 in unknown samples. This method ensures high sensitivity and specificity, making it ideal for research applications.
**Storage and Handling:**
- Store all reagents at 2–8°C.
- Samples should be stored at -20°C and avoid repeated freeze-thaw cycles.
- Centrifuge cell culture supernatants, tissue homogenates, and other biological fluids to remove particulates before use.
- Avoid hemolysis and granules in samples.
**Materials Required but Not Supplied:**
- 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:**
- 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 solutions A and B (6.0 ml each)
- Stop solution (6.0 ml)
- Closure plate membrane (2 pieces)
- User manual and sealed bags
**Preparation Instructions:**
- Dilute 1 volume of 20X Wash solution with 19 volumes of deionized or distilled water to make 1X solution.
- Allow all reagents to reach room temperature (20–25°C) before use.
- Do not thaw samples or reagents using water baths.
- Use only the reagents provided by the manufacturer.
- Do not use reagents beyond their expiration date.
- Always use fresh pipette tips to prevent cross-contamination.
**Assay Procedure:**
1. Prepare all reagents and standards before starting the assay.
2. Add 50 μL of standard or sample to the appropriate wells (duplicate).
3. Add 100 μL of HRP-Conjugate reagent to each well except the blank.
4. Incubate for 60 minutes at 37°C.
5. Wash the microtiter plate 4 times with 1X Wash solution.
6. Add 50 μL of Chromogen A and 50 μL of Chromogen B to each well.
7. Incubate for 15 minutes at 37°C, protecting from light.
8. Add 50 μL of Stop Solution to each well.
9. Measure OD at 450 nm using a microplate reader.
**Data Interpretation:**
- Plot the average OD values of the standards against their concentrations to generate a standard curve.
- Calculate the mean OD for each sample and subtract the blank value.
- Determine the FGF-9 concentration by comparing the sample OD to the standard curve.
**Performance Characteristics:**
- Intra-assay CV <15%, Inter-assay CV <15%
- Assay range: 15.6 pg/mL to 500 pg/mL
- Sensitivity: <10 pg/mL
- Cross-reactivity: No significant cross-reaction with other proteins
**Storage:**
- Store unused strips at 2–8°C in the desiccant-sealed pouch.
- Long-term storage: -20°C for up to 6 months.
**Safety Notes:**
- Handle all samples as potentially hazardous.
- Wear gloves during the procedure.
- Discard waste after 30 minutes of inactivation.
- Avoid contact with skin or eyes.
- Keep chromogen B away from heat and flame due to its acetone content.
This kit provides a reliable and efficient method for FGF-9 detection in research settings, ensuring consistent and reproducible results. Always follow the instructions carefully to maintain accuracy and safety.
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