Testing Principle：
This kit uses double antibody sandwich ELISA.Specific anti-human TNF-α Capture Antibody is precoated on a high affinity conjugate plate.  Standard substance, sample to be tested and biotin labeled detection antibody were added successively into the plate well of the enzyme label. After fully shaking and mixing, the samples were placed at room temperature for a 2-hour incubation process. The TNF-α in the samples was bound to the solid phase antibody and detection antibody.  After washing thoroughly to remove free and unbound ingredients, horseradish peroxidase labeled streptavidin-HRP (SA-HRP) is added.  After washing, signal enhancer was added for incubation. After washing and removing unbound substances, SA-HRP was added again.  After washing again, TMB color substrate was added and incubated in dark at room temperature for color development.  The depth of color reaction was positively correlated with the concentration of TNF-α in the sample.  The termination solution was added to terminate the reaction, and the absorbance value was measured at 450 nm detection wavelength (correction wavelength 570-630 nm) using a microplate analyzer.  

Detection type：Double sandwich enzyme immunoassay

Form: pre-coated 96-well plate

Sample type: Cell culture supernatant, Serum, Plasma

Sample quantity：100 μl

Kit composition：pre-coated 96-well plate,Standards,Standard, TNF-α Detector Antibody, Standard Diluent, Detector Buffer, Signal Enhancer, Signal Enhancer Diluent, TMB Color Substrate, Washing Solution, Termination Solution, SA-HRP, Sealing Plate Mask and Manual  

Sensitivity：0.16 pg/mL

Detection range：1.56-100 pg/mL

Recovery rate：100-108%

Storage：2-8℃

The standard curve：

Background：

Tumor necrosis factor α(TNF-α), also known as cachexin and TNFSF1A, is an adipokine that is involved in systemic inflammation and is one of the cytokines that stimulate acute phase responses. It is produced mainly by activated macrophages, but can also be secreted by other types of cells, such as CD4+ lymphocytes, NK cells, neutrophils, mast cells, eosinophils, and neurons. It has a variety of regulatory functions in the immune response, but also can be used as a potential pyrogen. In response to stimuli (infectious agents or tissue damage), TNF-α circulates throughout the body, activates neutrophils, alters the properties of vascular endothelial cells, regulates metabolic activity in other tissues, and exhibits tumor-killing activity by inducing local coagulation. TNF-α plays an important role in the pathogenesis of inflammation in joint tissues and other tissues. Recently, more and more information indicates that TNF-α is also involved in the pathogenesis of AIDS. The determination of TNF-α is also useful for transplantation studies.