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

Core Product Specifications and Parameters

Biological Background: TNF-α Function and Localization

• Tumor necrosis factor (TNF-α), also known as cachectin and TNFSF2, is a 26 kDa cytokine encoded by the TNF gene on chromosome 6p21.3.

• Primarily secreted by macrophages, TNF-α functions as a potent endogenous pyrogen, inducing fever directly and via IL-1, and is a key mediator of cachexia in chronic inflammation and cancer.

• It binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR, triggering signaling cascades that regulate cell survival, apoptosis, and differentiation across a broad range of cell types.

• The cytokine is synthesized as a type II transmembrane protein; its soluble ectodomain is shed by TACE/ADAM17, enabling both local juxtacrine and systemic endocrine-like activities.

• Post-translational modifications include glycosylation, disulfide bond formation, myristoylation, and phosphorylation, which are critical for proper folding, trafficking, and receptor binding.

• In rheumatoid arthritis, TNF-α promotes autoimmune inflammation by impairing regulatory T-cell (Treg) function through PP1-mediated dephosphorylation and inactivation of the FOXP3 transcription factor (PubMed:23396208).

  Related references: PMID:23396208

• TNF-α contributes to anticancer immunity by mediating neutrophil-dependent cytotoxicity against bladder cancer cells in combination with SMAC mimetic drugs (PubMed:16829952, PubMed:22517918).

  Related references: PMID:16829952, PMID:22517918

• It synergistically induces VEGF production with IL-1β and IL-6 to promote angiogenesis (PubMed:12794819) and stimulates osteoclastogenesis, driving bone destruction in inflammatory diseases (By similarity).

  Related references: PMID:12794819

Experimental Guidance and Technical Tips

• Since the immunogen spans the extracellular domain (amino acids 77-233), the antibody is expected to recognize both the membrane-bound and soluble forms of TNF-α.
• For intracellular flow cytometry, ensure proper permeabilization and fixation protocols to allow access to cytoplasmic TNF-α; consider using a known positive control such as activated monocytes.
• Validate the antibody in the relevant sample system; human whole blood or PBMCs stimulated with LPS can be used to induce TNF-α expression for intracellular staining.
• Because TNF-α is secreted, a protein transport inhibitor (e.g., brefeldin A or monensin) is recommended during cell stimulation to accumulate intracellular protein.
• Take care to avoid non-specific binding by titrating the antibody and using appropriate isotype controls.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio provides diagnostic manufacturers, labs, and research institutes with one-stop access to IVD raw materials, from concept to clinic. For TNF-α-related immuno-oncology and inflammation research, we supply validated antibody pairs, optimized monoclonal/polyclonal antibodies, bulk ancillary reagents, and can source rare targets to support your assay development. Contact us to streamline your procurement and accelerate your project timeline.