Mouse TNF alpha ELISA Kit

• Microplate reader with 450 nm wavelength filter
• Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
• Incubator
• Deionized or distilled water
• Absorbent paper
• Buffer resevoir

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Tumor Necrosis Factor-alpha (TNF-α) is a cytokine that plays a pivotal role in immune responses and inflammation. It is a pro-inflammatory protein involved in various physiological and pathological processes, including host defense, tissue repair, and the pathogenesis of autoimmune diseases. TNF-α is produced by multiple cell types, such as macrophages and T cells, and exerts its effects by binding to two distinct receptors, TNFR1 and TNFR2. Upon activation, TNF-α triggers signaling pathways that regulate gene expression, immune cell recruitment, and cellular responses. Due to its critical role in inflammatory conditions, TNF-α has been extensively studied, leading to the development of therapeutic agents targeting this cytokine to modulate immune responses and alleviate inflammatory diseases.

The TNF-alpha gene in mice, also known as Tnf, is located on chromosome 17. It encodes a pro-inflammatory cytokine protein, TNF-α, which is involved in multiple cellular processes, including immune responses, apoptosis, and inflammation. The Tnf gene consists of four exons and three introns and undergoes transcriptional regulation to produce TNF-α mRNA.

TNF-α plays a crucial role in regulating immune responses and inflammation. It acts as a potent pro-inflammatory cytokine, initiating and amplifying the immune response in various physiological and pathological conditions. TNF-α is produced by several cell types, including macrophages, T cells, and natural killer cells. It can induce the expression of other cytokines, chemokines, and adhesion molecules, promoting leukocyte recruitment, tissue repair, and host defense mechanisms.

TNF-alpha (TNF-α) exerts its biological effects by binding to two distinct cell surface receptors: TNF receptor 1 (TNFR1) and TNF receptor 2 (TNFR2). These receptors belong to the TNF receptor superfamily, which is characterized by the presence of cysteine-rich repeats in their extracellular domains. While both TNFR1 and TNFR2 can bind TNF-α, they differ in their downstream signaling and biological functions.The balance between TNFR1 and TNFR2 signaling is critical in determining the overall cellular response to TNF-α.

1. TNF Receptor 1 (TNFR1): Also known as p55 or CD120a, TNFR1 is widely expressed on various cell types, including immune cells, endothelial cells, and many other cell types. TNFR1 signaling can activate multiple pathways, including the canonical NF-κB pathway, which leads to the transcriptional activation of genes involved in inflammation, cell survival, and immune responses. TNFR1 activation can also initiate apoptotic pathways, promoting cell death under certain conditions.

2. TNF Receptor 2 (TNFR2): Also known as p75 or CD120b, TNFR2 is expressed on a more restricted set of cells, such as immune cells, endothelial cells, and some neuronal cells. TNFR2 signaling can activate similar pathways to TNFR1, including the NF-κB pathway. Additionally, TNFR2 can promote cell proliferation, survival, and immune cell activation. It plays a crucial role in regulating immune responses and tissue homeostasis.

The dysregulation of TNF-α has been implicated in various human diseases, including autoimmune disorders, chronic inflammation, and cancer. Elevated levels of TNF-α have been observed in conditions such as rheumatoid arthritis, Crohn's disease, and psoriasis. Conversely, decreased TNF-α production or impaired signaling can lead to immunodeficiency and increased susceptibility to infections. Due to its clinical significance, TNF-α has become an important therapeutic target for the development of drugs and therapies.

Given the crucial role of TNF-α in inflammation and immune regulation, researchers have developed both agonists and antagonists/blockers to modulate its activity. TNF-α agonists, such as etanercept and adalimumab, are engineered proteins that mimic the natural ligand, enhancing TNF-α signaling. These agonists have been employed in the treatment of certain autoimmune diseases. Conversely, TNF-α antagonists/blockers, including infliximab and certolizumab, bind to TNF-α and prevent its interaction with receptors, thereby reducing the inflammatory response. These antagonists have shown efficacy in treating various inflammatory conditions, such as rheumatoid arthritis and inflammatory bowel disease.