Detection of TNF‑ alpha in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs) treated with PMA and Ca2+ ionomycin were stained with Mouse Anti-Human TNF‑ alpha PE‑conjugated Monoclonal Antibody (Catalog # IC210P, filled histogram) or isotype control antibody (Catalog # IC002P, open histogram). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.
Preparation and Storage
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Protect from light. Do not freeze.
12 months from date of receipt, 2 to 8 °C as supplied.
TNF-alpha, designated TNFSF2, is a trimeric glycoprotein active in both membrane bound and secreted forms. TNF-alpha is produced by several lymphoid cells as well as by astrocytes, endothelial cells, and smooth muscle cells. TNF-alpha binds to TNF RI and TNF RII present on virtually all cell types where it triggers the activation of multiple signal transduction pathways and modulates the expression of a wide variety of genes.
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but also provides information about sample types, species, and experimental conditions.
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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