Human TNF-α ELISA Kit

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Cat: abs510006

Size: 96T

Human TNF-α ELISA Kit

Testing Principle

The quantitative sandwich enzyme immunoassay technique is employed in this kit. The high affinity plate is pre-coated with a specific anti-human TNF-α antibody. The wells of the enzyme-labeled plate are then filled with standards, test samples, and biotinylated detection antibodies. Following an incubation period, the present TNF-α in the samples binds with the solid-phase antibodies and detection antibodies, forming immune complexes. Unbound material is washed away, and horseradish peroxidase-labeled streptavidin (HRP) is added. Subsequently, a chromogenic substrate is introduced, and the color development occurs in darkness. The reaction is halted with a stop solution, and the absorbance is measured at 450 nm wavelength (with reference correction at 540 nm or 570 nm).

Detection type: sandwich enzyme immunoassay

Form: pre-coated 96-well plate

Sample type: Cell culture supernatant, Serum, Plasma

Sample quantity: 100ul

Kit composition:

This microplate kit includes a 96 well polysyrene plate with a capture antibody already coated onto it, along with a standard and detection antibody. Also included are a 10×reagent diluent, Color reagent (A and B), 25×wash buffer, and Stop solution to complete the ELISA assay. Additionally, the kit includes ELISA plate sealers and a specification for reference.

Sensitivity:  0.68 pg / mL

Detection range:  15.6 - 1000 pg/mL

Recovery rate: 87-104%

Storage: 2-8 ° C

The standard curve:

Background: TNF-α 

TNF-α, also referred to as TNFSF1A or cachectin, is a prototypical ligand in the tumor necrosis factor superfamily with pleiotropic effects. The cytokine is heavily involved in several critical processes in the human body, such as lipid metabolism, immune system development, apoptosis, and inflammatory response.
Moreover, TNF-α appears to play essential roles in the pathogenesis of several diseases, including autoimmune disorders, obesity, septic shock, asthmatic conditions, neuropathic pain, Crohn's disease, rheumatoid arthritis, and many forms of cancer. Its significance in these health conditions makes it a vital target of clinical research with the hope that the findings will help develop novel therapies and interventions.

The type II transmembrane protein, human TNF-α, has a molecular weight of 26 kDa. It comprises of an intracellular domain of 35 amino acids, a transmembrane segment of 21 amino acids, and an extracellular domain (ECD) of 177 amino acids. The ECD region of human TNF-α has a high degree of homology with other animals such as rhesus monkey, cow, dog, cotton mouse, horse, cat, mouse, pig, and rat. The protein can be expressed by various cell types including immune cells, epithelial cells, endothelial cells, and tumor cells. TNF-α is capable of inducing tumor cell lysis and the destruction of virus-infected cells. It binds to soluble TNF RI to produce transduction signals in downstream cells. TACE/ADAM17 triggers the shedding of cell membranes containing TNF-α, leading to the release of the active cytokine, which is a trimeric structure consisting of a soluble extracellular form of TNF-α, weighing about 55 kDa.

TNF-α has two receptors,which are TNF RI and TNF RII.The molecular weight of TNF RI is 55-60 kDa and it is widely expressed.TNF RII has a molecular weight of 78-80 kDa and is just expressed in hematopoietic cells. Both of them are expressed as homologous trimer. TNF-α promotes NFκB activation with a similar binding affinity to TNF RI and TNF RII. Both the two soluble receptors are released into human serum and urine and they can neutralize TNF-α activity. However, only TNF RI has a cell death domain that can induce apoptosis.

Technical hints: For research use only, not for in vitro diagnosis.

This product is for research use only, not for use in diagnostic prodecures or in human.

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