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Horse Tumor necrosis factor α, TNF-α ELISA Kit

Catalog #BYEK3209
In stockyes
Datasheet MSDS

Catalog No.: BYEK3209

Size: 96T

Range: 4.375 ng/L- 70 ng/L 

Sensitivity:0.1 ng/L

Storage and Expiration: Store at 2-8℃ for 6 months. 

Application: For quantitative detection of TNF-α in horse serum, plasma, tissue homogenate or cell culture supernatant.

Tumor necrosis factor, formerly known as tumor necrosis factor-alpha or TNF-α, is a cytokine involved in systemic inflammation and is a member of a group of cytokines that stimulate the acute phase reaction. It was originally identified in Human serum after injection with Mycobacterium bovis strain bacillus Calmette-Guerin (BCG) and endotoxin. TNF is primarily produced as a 212-amino acid-long type II transmembrane protein arranged in stable homotrimers. The human TNF gene maps to chromosome 6p21.3, spans about 3 kilobases and contains 4 exons. The primary role of TNF is in the regulation of immune cells. Dysregulation of TNF production has been implicated in a variety of human diseases, including Alzheimer's disease, cancer, major depression, and inflammatory bowel disease (IBD). 

Principle of the Assay  
This kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. The purified anti- TNF-α antibody was pre-coated onto 96-well plates. And the HRP conjugated anti- TNF-α antibody was used as detection antibodies. The standards, test samples and HRP conjugated detection antibody were added to the wells subsequently, mixed and incubated, then, unbound conjugates were washed away with wash buffer. TMB substrates (A & B) 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 TNF-α amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of TNF-α can be calculated.

Kit components
1. One 96-well plate pre-coated with anti- horse TNF-α antibody
2. Standard: 0.3 ml × 6 tubes (0 ng/L, 4.375 ng/L, 8.75 ng/L, 17.5 ng/L, 35 ng/L, 70 ng/L)
3. Wash buffer (20×): 25 ml. Dilution: 1:20
4. Sample diluent buffer: 6 ml
5. HRP conjugated anti- horse TNF-α antibody (RTU): 10 ml
6. Stop solution: 6 ml
7. TMB substrate A:  6ml
8. TMB substrate B:  6ml
9. Plate sealer:   2
10. Hermetic bag:  1

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. Absorbent filter papers
8. Plastic or glass container with volume of above 1L

Other Information

Please check datasheet for protocol.