Human TNF-alpha Fluorescein-conjugated Antibody Summary
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 PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs) treated with PMA and Ca2+ionomycin for 3.5 hours were stained with (A) Mouse Anti-Human TNF-alpha CFS-conjugated Monoclonal Antibody (Catalog # IC210F) or (B) isotype control antibody (IC002F) and Mouse anti-Human CD3 APC-conjugated Monoclonal Antibody (FAB100A). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (FC005). Staining was performed using our Staining Intracellular Molecules 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
Citations for Human TNF-alpha Fluorescein-conjugated Antibody
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 4
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Monocyte Differentiation towards Protumor Activity Does Not Correlate with M1 or M2 Phenotypes
Authors: G Karina Chimal-Ram
J Immunol Res, 2016-06-08;2016(0):6031486.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry -
Expression of the T cell receptor alpha/beta on a CD123+ BDCA2+ HLA-DR+ subpopulation in head and neck squamous cell carcinoma.
Authors: Thiel A, Kesselring R, Pries R, Puzik A, Wittkopf N, Wollenberg B
PLoS ONE, 2011-01-11;6(1):e15997.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry -
Kinin receptor expression during Staphylococcus aureus infection.
Authors: Bengtson SH, Phagoo SB, Norrby-Teglund A, Pahlman L, Morgelin M, Zuraw BL, Leeb-Lundberg LM, Herwald H
Blood, 2006-05-30;108(6):2055-63.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry -
Nucleoporation of dendritic cells: efficient gene transfer by electroporation into human monocyte-derived dendritic cells.
Authors: Lenz P, Bacot SM, Frazier-Jessen MR, Feldman GM
FEBS Lett., 2003-03-13;538(1):149-54.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry
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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Would you expect FAB210 and IC210 to give different intracellular staining results and why?
Yes. 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 thier receptor and membrane bound forms. IC210 on the other hand detects the cytoplasmic form o fthe TNF-alpha, that is cells that are actively secreting TNF-alpha intracellularly. If FAB210 is used for intracellular staining, there is a chance that antibody will report addivitve staining of cell surface TNF-alpha as well as intracellular TNF-alpha upon permeabilization.
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