Human TNF-alpha Membrane Form PE-conjugated Antibody Summary
Gly57-Leu233 (predicted)
Accession # P01375
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
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Detection of TNF‑ alpha in Human PBMC Monocytes by Flow Cytometry. Human PBMCs were treated with 1μg/ml LPS for 4 hours then stained with (A) Mouse Anti-Human TNF-a Membrane Form PE-conjugated Monoclonal Antibody (Catalog # FAB210P) or (B) isotype control antibody (IC002P) followed by Mouse anti-Human CD14 APC-conjugated Monoclonal Antibody (FAB3832A). Staining was perfomed using our Staining Membrane-associated Proteins protocol.
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Preparation and Storage
- 12 months from date of receipt, 2 to 8 °C as supplied.
Background: TNF-alpha
Tumor necrosis factor alpha (TNF-alpha, TNF- alpha, TNFA ), also known as Cachectin and TNFSF2, is the prototypic ligand of the TNF superfamily. It is a pleiotropic molecule that plays a central role in inflammation, immune system development, apoptosis, and lipid metabolism. TNF- is produced by several lymphoid cells as well as by astrocytes, endothelial cells, and smooth muscle cells. Human TNF-alpha consisits of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 177 aa extracellular domain (ECD). Within the ECD, human TNF-alpha shares 97% aa sequence identity with rhesus and 71%-92% with bovine, canine, cotton rat, equine, feline, mouse, porcine, and rat TNF-alpha. TNF-alpha is produced by a wide variety of immune, epithelial, endothelial, and tumor cells. TNF-alpha is assembled intracellularly to form a noncovalently linked homotrimer which is expressed on the cell surface. Cell surface TNF-alpha can induce the lysis of neighboring tumor cells and virus infected cells, and it can generate its own downstream cell signaling following ligation by soluble TNFR I. Shedding of membrane bound TNF-alpha by TACE/ADAM17 releases the bioactive cytokine, a 55 kDa molecular weight soluble trimer of the TNF-alpha extracellular domain. TNF-alpha binds the ubiquitous 55-60 kDa TNF RI and the hematopoietic cell-restricted 80 kDa TNF RII, both of which are also expressed as homotrimers present on virtually all cell types. Both type I and type II receptors bind TNF-alpha with comparable affinity, although only TNF RI contains a cytoplasmic death domain which triggers the activation of apoptosis. Soluble forms of both types of receptors are released and can neutralize the biological activity of TNF-alpha.
Product Datasheets
FAQs
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Why do FAB210 and IC210 give different intracellular staining results?
FAB210 was developed to detect cells that are expressing cell surface TNF-alpha and the antibody is able to recognize cells that have bound TNF-alpha on their surface through their receptor and membrane bound forms. The IC210 on the other hand detects the cytoplasmic form of TNF-alpha, that is cells which are actively secreting TNF-alpha intracellularly. If FAB210 is used for intracellular staining, there is a chance that antibody will report additive staining of cell surface TNF-alpha as well as intracellular TNF-alpha upon permeabilization.
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