Rat IL-3 ELISA Kit

Catalog No.: BEK1284

Size: 96T

Range: 7.8 pg/ml - 500 pg/ml

Sensitivity < 3 pg/ml

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

Application: For quantitative detection of IL-3 in rat serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Interleukin 3 (IL-3) is a hematopoietic colony-stimulating factor that is capable of supporting the proliferation of a broad range of hematopoietic cell types. It is chemically synthesized by means of an automated peptide synthesizer and is shown to have the biological activities attributed to native IL-3. It is secreted by basophils and activated T cells to support growth and differentiation of T cells from the bone marrow in an immune response. It can improve the body's natural response to disease as part of the immune system. It acts by binding to the interleukin-3 receptor. IL3 infusion expands an early cell population that subsequently requires the action of a later-acting factor such as GMCSF to complete its development.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-IL-3 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-IL-3 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-3 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of IL-3 can be calculated.

Kit components

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