Rat IFNγ ELISA Kit

Catalog No.: BEK1283

Size: 96T

Range: 31.2 pg/ml - 2000 pg/ml

Sensitivity < 6 pg/ml

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

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

Introduction

Interferon-gamma (IFN-γ), early in its history was known as immune interferon, was recognized in 1970. It dimerized soluble cytokine and is the only member of the type II class of interferons. The IFN-γ monomer consists of a core of six α-helices and an extended unfolded sequence in the C-terminal region. It is produced predominantly by natural killer (NK) and natural killer T (NKT) cells. It is critical for innate and adaptive immunity against viral and intracellular bacterial infections and for tumor control. Interferon-γ 1b is used to treat chronic granulomatous disease and osteopetrosis.

Principle of the Assay  

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

Kit components

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