Mouse TNF Alpha ELISA Kit

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Cat: abs520010

Size: 96T

Mouse TNF alpha ELISA Kit

Testing Principle

The sandwich enzyme immunoassay technique is employed in this kit. The high affinity plate is pre-coated with a specific anti-mouse TNF-α antibody. Subsequently, standards, test samples, and biotinylated detection antibodies are introduced to the wells of the plate that has been labeled with enzymes. Following an incubation period, the TNF-α in the samples interacts with the solid-phase antibodies and the detection antibodies, resulting in the formation of immune complexes. After removing any unbound material through washing, horseradish peroxidase-labeled streptavidin (HRP) is added. Another round of washing is carried out before introducing a chromogenic substrate, which initiates the development of color in a dark environment. The reaction is halted with the addition of a stop solution, and the absorbance is measured at a wavelength of 450 nm (reference correction wavelength of 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 package includes a 96 well polysyrene microplate that has been coated with a capture antibody. Additionally, a standard, detection antibody, and 10×reagent diluent are included. Other components of the kit include Color reagent A and B, 25×wash buffer, and Stop solution. The package also includes ELISA plate sealers and a specification.

Sensitivity:  5.5 pg/mL

Detection range:  31.2 - 2,000 pg/mL

Recovery rate: 88-107%

Storage: 2-8 ° C

The standard curve:

Background: TNF-α 

TNF-α, or cachectin and TNFSF1A, is a significant molecule belonging to the tumor necrosis factor superfamily. Its functions are diverse, ranging from participating in the inflammatory response, immune system development, apoptosis, to lipid metabolism regulation. Pathologically, TNF-α is implicated in various conditions such as asthma, Crohn's disease, rheumatoid arthritis, neuropathic pain, obesity, type 2 diabetes, septic shock, autoimmunity, and cancer. It serves as a crucial factor in these processes and understanding its role is of great significance. The impact of TNF-α extends beyond a single disease entity, encompassing a wide range of conditions where its dysregulation plays a pivotal role in disease progression.

The human TNF-α is a transmembrane protein with a molecular weight of 26 kDa, featuring 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 shares 97% amino acid homology with rhesus monkey and 71-92% homology with other animals such as cow, dog, mouse, pig, and rat. Cells that express TNF-α include immune cells, tumor cells, endothelial cells, and epithelial cells. TNF-α can bind to the soluble TNF RI to activate transmembrane signals in downstream cells and lyse tumor and virus-infected cells. TACE/ADAM17 induces the shedding of cell membranes containing TNF-α to release the active TNF-α cytokine, which comprises a soluble trimeric structure with a molecular weight of 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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