Rat IL-5 ELISA Kit

Catalog No.: BEK1285

Size: 96T

Range: 15.6 pg/ml -1000 pg/ml

Sensitivity < 10 pg/ml

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

Application: For quantitative detection of IL-5 in rat serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Interleukin 5 (IL-5) is an interleukin produced by T helper-2 cells and mast cells. It is a 115-amino acid-long TH2 cytokine that is part of the hematopoietic family. It is expressed by eosinophils and has been observed in the mast cells of asthmatic airways by immunohistochemistry. IL-5 expression is regulated by several transcription factors including GATA3. IL-5 is a key mediator in eosinophil activation. It has long been associated with the cause of several allergic diseases including allergic rhinitis and asthma, where in a large increase in the number of circulating, airway tissue, and induced sputum eosinophils have been observed. Given the high concordance of eosinophils and, in particular, allergic asthma pathology, it has been widely speculated that eosinophils have an important role in the pathology of this disease.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- IL-5 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- IL-5 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-5 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of IL-5 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-rat IL-5 antibody
2. Lyophilized rat IL-5 standards: 2 tubes (10 ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-rat IL-5 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