Rabbit anti-Human TNF-α Monoclonal Antibody for Intracellular Flow Cytometry - P01375

Core Product Specifications and Parameters

Biological Background: Tumor necrosis factor Function and Localization

• Tumor necrosis factor alpha (TNF-α), also known as cachectin or TNFSF2, is a key cytokine of the TNF superfamily and exists as both a transmembrane and soluble form. The soluble form is derived from proteolytic cleavage of the membrane-bound precursor.
• TNF-α binds to its receptors TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR, triggering signaling pathways that lead to apoptosis, cell survival, inflammation, and immune modulation. It is a potent pyrogen, causing fever directly or via IL-1 induction, and is involved in cachexia induction.
• In rheumatoid arthritis, TNF-α impairs regulatory T-cell function by up-regulating protein phosphatase 1 (PP1), which dephosphorylates FOXP3 at Ser-418, thereby inactivating FOXP3 and rendering Treg cells functionally defective. Related references: PMID:23396208
• TNF-α acts as a mediator of cell death in anticancer immune responses, exemplified by its role with BCG-stimulated neutrophils and DIABLO/SMAC mimetics in bladder cancer cells. Related references: PMID:16829952, PMID:22517918, PMID:23396208
• It promotes angiogenesis by synergistically inducing VEGF production with IL1B and IL6. Related references: PMID:12794819
• TNF-α induces insulin resistance in adipocytes by inhibiting insulin-induced IRS1 tyrosine phosphorylation and glucose uptake, partly via GKAP42 protein degradation, and promotes osteoclastogenesis, contributing to bone resorption.
• Subcellularly, TNF-α is located at the cell membrane, in intracellular membranes, and is secreted. The TNF intracellular domain (ICD) form can additionally induce IL12 production in dendritic cells.
• Post-translational modifications include disulfide bond formation, glycosylation, myristoylation, and phosphorylation, which are critical for its stability and activity.

Experimental Guidance and Technical Tips

• The immunogen (amino acids 77-233) corresponds to the mature soluble form of TNF-α, lacking the N-terminal transmembrane domain. This design may favor detection of both secreted and intracellular precursor forms.
• The conjugate is optimized for flow cytometry; for intracellular staining, consider using a fixation/permeabilization protocol validated for cytokines. Include appropriate isotype controls and confirm staining in stimulated vs. unstimulated samples.
• Perform titration experiments to determine the optimal antibody concentration for your specific experimental setup, as staining indices can vary across cell types and activation states.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio offers diagnostic manufacturers and researchers streamlined access to high-quality IVD raw materials, including validated antibody pairs, optimized monoclonal antibodies, and bulk ancillary reagents. For TNF-α and other cytokine targets, we provide comprehensive sourcing support from concept to clinic, enabling efficient development of immunoassays and cellular analysis tools. Whether you need rare target reagents or established catalog products, our one-stop platform bridges your procurement needs with reliable supply solutions.