Human Liver Fatty acid Binding Protein (L-FABP) ELISA Kit

The Human Liver Fatty Acid Binding Protein (L-FABP) ELISA Kit is a highly specific and sensitive assay designed for the quantification of L-FABP levels in human serum, plasma, culture media, or any biological fluid. The ELISA (Enzyme-Linked Immunosorbent Assay) kit uses a sandwich-ELISA method and provides a reliable solution for L-FABP detection across various research applications in life sciences, biomedical studies, and clinical research.

Principle of the Assay

This sandwich-ELISA method involves the following steps:

1. The Microelisa strip plate is pre-coated with an antibody specific to L-FABP.
2. Standards or samples are added to the plate, where they bind to the antibody.
3. An HRP-conjugated antibody specific for L-FABP is added, followed by incubation to allow the formation of an antigen-antibody complex.
4. The unbound components are washed away, and the TMB substrate solution is added to the wells.
5. L-FABP presence is detected by the change in color from blue to yellow after the addition of the stop solution.
6. The optical density (OD) is measured at 450 nm, and the concentration of L-FABP is calculated using a standard curve.

Key Features and Benefits

• High Sensitivity and Precision:
  This kit provides a sensitive assay with a detection limit as low as 2.2 pg/mL. It ensures precise quantification of L-FABP levels across a wide range of samples.
• Wide Detection Range:
  The assay can detect L-FABP concentrations from 10 pg/mL to 800 pg/mL, making it suitable for both low and high concentration samples.
• Convenient Ready-to-Use Kit:
  The Human L-FABP ELISA kit comes with all the necessary components, including standard diluents, HRP-conjugated reagents, and chromogen solutions, ensuring easy setup and minimal preparation time.
• Compatible with Multiple Sample Types:
  This ELISA kit can be used with various biological fluids such as serum, plasma, urine, culture media, and cell culture supernatants, providing flexibility for different experimental setups.
• Precision and Reproducibility:
  The intra-assay precision (CV < 10%) and inter-assay precision (CV < 12%) ensure reliable and reproducible results for quantifying L-FABP in samples.
• Simple and Fast Procedure:
  The ELISA protocol is straightforward, with easy incubation steps and minimal washing, reducing the time required for each assay and enabling high-throughput applications.
• Cost-Effective:
  The kit’s bulk packaging provides an economical solution for labs with high sample volumes, making it ideal for large-scale studies.

Materials Provided in the Kit

Sample Preparation

To ensure accurate and reliable results, follow these sample preparation steps:

1. Serum:
   • Collect blood and allow it to clot at room temperature for 10-20 minutes.
   • Centrifuge at 2,000-3,000 rpm for 20 minutes. If precipitates form, centrifuge again.
2. Plasma:
   • Collect whole blood with an anticoagulant (e.g., EDTA or citrate).
   • Incubate at room temperature for 10-20 minutes, then centrifuge at 2,000-3,000 rpm for 20 minutes.
   • Collect plasma as the supernatant.
3. Urine Samples:
   • Collect urine in aseptic tubes, centrifuge at 2,000-3,000 rpm for 20 minutes, and collect the supernatant.
4. Cell Culture Supernatants:
   • Collect the culture medium from cell cultures, centrifuge, and transfer the supernatant into clean tubes.
5. Tissue Samples:
   • Homogenize frozen tissue with PBS, centrifuge, and collect the supernatant for analysis.

Assay Procedure

1. Dilute Standards:
   • Prepare a standard curve by serially diluting the standard in microplate wells.
   • Pipette 50 μL of each dilution into the respective wells.
2. Add Samples:
   • Add 40 μL of sample dilution buffer and 10 μL of sample to each well. Mix gently.
3. Incubate:
   • Incubate the plate at 37°C for 30 minutes, then wash to remove unbound components.
4. Add HRP-Conjugate:
   • Add 50 μL of HRP-conjugate to each well and incubate for another 30 minutes.
5. Wash:
   • Wash the plate 5 times to remove excess reagent.
6. Add Chromogen Solution:
   • Add 50 μL of Chromogen Solution A and 50 μL of Chromogen Solution B to each well, then incubate for 15 minutes at 37°C, avoiding light.
7. Terminate Reaction:
   • Add 50 μL of Stop Solution to each well. The color will change from blue to yellow.
8. Read OD:
   • Measure the optical density (OD) at 450 nm using a microplate reader.

Calculating Results

1. Plot the Standard Curve:
   Plot the OD values of the standard on the Y-axis and the corresponding concentrations on the X-axis.
2. Sample Concentration:
   Compare the OD of each sample to the standard curve to determine the L-FABP concentration in the sample.
3. Dilution Factor:
   If a sample was diluted before testing, multiply the measured concentration by the dilution factor to obtain the original concentration.

Precision and Sensitivity

• Intra-Assay Precision (CV < 10%)
• Inter-Assay Precision (CV < 12%)
• Sensitivity: 2.2 pg/mL
• Assay Range: 10 pg/mL to 800 pg/mL

Storage and Validity

• Storage: Store the kit at 2-8°C upon receipt.
• Shelf Life: The kit is valid for 6 months when stored properly.

The Human Liver Fatty Acid Binding Protein (L-FABP) ELISA Kit provides a highly sensitive and precise method for measuring L-FABP levels in human serum, plasma, and culture media. With reliable results, high reproducibility, and a straightforward procedure, this kit is an invaluable tool for biomedical research, clinical studies, and medical applications. Its flexibility and user-friendly design make it suitable for a wide range of research and diagnostic applications.