Human IL-33 ELISA Kit

Catalog No.: BEK1314

Size: 96T

Range: 15.6 pg/ml - 1000 pg/ml

Sensitivity < 2 pg/ml

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

Application: For quantitative detection of IL-33 in human serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Interleukin 33 (IL-33) is a protein that in humans is encoded by the IL33 gene. Interleukin 33 is a member of the IL-1 family that potently drives production of T helper-2 (Th2)-associated cytokines (e.g., IL-4). IL33 is a ligand for ST2 (IL1RL1), an IL-1 family receptor that is highly expressed on Th2 cells, mast cells and group 2 innate lymphocytes. IL-33 is expressed by a wide variety of cell types, including fibroblasts, mast cells, dendritic cells, macrophages, osteoblasts, endothelial cells, and epithelial cells.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- IL-33 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- IL-33 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-33 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of IL-33 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-human IL-33 antibody
2. Lyophilized human IL-33 standards: 2 tubes (10ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-human IL-33 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