Human TNF-alpha Fluorescein-conjugated Antibody

Catalog # Availability Size / Price Qty
IC210F
Human TNF-alpha  Fluorescein-conjugated Antibody in Intracellular Staining by Flow Cytometry
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Product Details
Citations (4)
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Human TNF-alpha Fluorescein-conjugated Antibody Summary

Species Reactivity
Human
Specificity
Detects cytoplasmic forms of human TNF-alpha.
Source
Monoclonal Mouse IgG1 Clone # 6402
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Recombinant human TNF-alpha
Formulation
Supplied in a saline solution containing BSA and Sodium Azide.
Label
Fluorescein

Applications

Recommended Concentration
Sample
Intracellular Staining by Flow Cytometry
10 µL/106 cells
See below

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.

Data Example

Intracellular Staining by Flow Cytometry Detection of TNF‑ alpha  in Human PBMCs by Flow Cytometry. View Larger

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 (filled histogram) or untreated (open histogram) were stained with Mouse Anti-Human TNF-a Fluorescein-conjugated Monoclonal Antibody (Catalog # IC210F) or isotype control antibody (Catalog # IC002F, not shown). 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.

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Preparation and Storage

Shipping
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.

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.

Long Name
Tumor Necrosis Factor alpha
Entrez Gene IDs
7124 (Human); 21926 (Mouse); 24835 (Rat); 397086 (Porcine); 280943 (Bovine); 403922 (Canine); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit)
Alternate Names
APC1 protein; Cachectin; Cachetin; DIF; TNF; TNF, monocyte-derived; TNFA; TNF-A; TNFalpha; TNF-alpha; TNF-alphacachectin; TNFATNF, macrophage-derived; TNFG1F; TNFSF1A; TNFSF2; TNFSF2TNF superfamily, member 2; tumor necrosis factor (TNF superfamily, member 2); tumor necrosis factor alpha; Tumor necrosis factor ligand superfamily member 2; tumor necrosis factor; tumor necrosis factor-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.

4 Citations: Showing 1 - 4
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  1. Monocyte Differentiation towards Protumor Activity Does Not Correlate with M1 or M2 Phenotypes
    Authors: G Karina Chimal-Ram
    J Immunol Res, 2016;2016(0):6031486.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  2. 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;6(1):e15997.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  3. 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;108(6):2055-63.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  4. 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;538(1):149-54.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry

FAQs

  1. 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.

  2. 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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