Mouse IL-12(P40) ELISA Kit

Catalog No.: BEK1286

Size: 96T

Range: 7.8 pg/ml - 500 pg/ml

Sensitivity < 2.7 pg/ml

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

Application: For quantitative detection of IL-12(P40) in mouse serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Interleukin 12 (IL-12) is an interleukin that is naturally produced by dendritic cells, macrophages and human B-lymphoblastoid cells (NC-37) in response to antigenic stimulation. IL-12 is composed of a bundle of four alpha helices. It is a heterodimeric cytokine encoded by two separate genes, IL-12A (p35) and IL-12B (p40). IL-12 plays an important role in the activities of natural killer cells and T lymphocytes. It mediates enhancement of the cytotoxic activity of NK cells and CD8+ cytotoxic T lymphocytes. IL-12 also has anti-angiogenic activity, which means it can block the formation of new blood vessels. IL12 p40 exists as a monomer and as a homodimer. The IL12B gene contains 8 exons, and maps to 5q31-q33.

Principle of the Assay

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- IL-12(P40) polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- IL-12(P40) 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-12(P40) amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of IL-12(P40) can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-mouse IL-12(P40) antibody
2. Lyophilized mouse IL-12(P40) standards: 2 tubes (10 ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-mouse IL-12(P40) 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
11. 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