Human IL-2RA / CD25 ELISA Kit

Catalog No.: BEK1325

Size: 96T

Range: 15.6 pg/ml - 1000 pg/ml

Sensitivity < 5 pg/ml

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

Application: For quantitative detection of IL-2RA / CD25 in human serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

The interleukin-2 receptor alpha chain (also called Tac antigen, P55, and mainly CD25) is a protein involved in the assembly of the high-affinity interleukin-2 receptor, consisting of alpha (IL2RA), beta (IL2RB) and the common gamma chain (IL2RG). As the name indicates, this receptor interacts with interleukin-2, a pleiotropic cytokine which plays an important role in immune homeostasis. The human protein interleukin-2 receptor subunit alpha is encoded by a gene called IL2RA with a length around 51,6 kb. Alternative names for this protein coding gene are IL2R, IDDM10 and TCGFR. Location of IL2RA in human genome is on the short arm of 10th chromosome (10p15.1).

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- IL-2RA / CD25 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- IL-2RA / CD25 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-2RA / CD25 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of IL-2RA / CD25 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-human IL-2RA / CD25 antibody
2. Lyophilized human IL-2RA / CD25 standards: 2 tubes (10 ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-human IL-2RA / CD25 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