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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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.
Have you used Human TNF-alpha PE-conjugated Antibody?
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