Human IL-18 ELISA Kit

Catalog No.: BEK1125

Size: 96T

Range: 62.5 pg/ml - 4000 pg/ml

Sensitivity < 8 pg/ml

Storage and Expiration: Store at 2-8℃ for 6 months, or at -20℃ for 12 months. 

Application: For quantitative detection of IL-18 in human serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Interleukin-18 (IL18), also known as interferon-gamma inducing factor, in humans is encoded by the IL18 gene, this gene contains 6 exons, and maps to 11q22.2-q22.3. It is a cytokine that belongs to the IL-1 superfamily and is produced by macrophages and other cells. It works by binding to the interleukin-18 receptor, and together with IL-12 it induces cell-mediated immunity following infection with microbial products like lipopolysaccharide (LPS). IL-18 has been implicated as an inflammatory mediator of Hashimoto's thyroiditis, the most common cause of autoimmune hypothyroidism. IL-18 is also able to induce severe inflammatory reactions, which suggests its role in certain inflammatory disorders.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- IL-18 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- IL-18 polyclonal antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the IL-18 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of IL-18 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-Human IL-18 antibody
2. Lyophilized Human IL-18 standards: 2 tubes (10 ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-Human IL-18 antibody (Concentrated): 130μl. Dilution: 1:100
5. Antibody diluent buffer: 12ml
6. Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130μl. Dilution: 1:100
7. ABC diluent buffer: 12ml
8. TMB substrate: 10ml
9. Stop solution: 10ml
10. Wash buffer (25X): 30ml

Note: Reconstitute standards and test samples with Kit Component 3.

Material Required But Not Provided

1. 37℃ incubator
2. Microplate reader (wavelength: 450nm)
3. Precise pipette and disposable pipette tips
4. Automated plate washer
5. ELISA shaker
6. 1.5ml of Eppendorf tubes
7. Plate cover
8. Absorbent filter papers
9. Plastic or glass container with volume of above 1L