Human TNF-alpha Fluorescein-conjugated Antibody

(4 citations)   
  • 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 Examples
Intracellular Staining by Flow Cytometry
Detection of TNF‑ alpha in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs) treated with PMA and Ca2+ ionomycin for 24 hours were stained with Mouse Anti-Human TNF‑ alpha Fluorescein‑conjugated Monoclonal Antibody (Catalog # IC210F, filled histogram) or isotype control antibody (Catalog # IC002F, 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
  • 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
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.
  • 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; 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
Related Research Areas
Citations:

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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Species
Applications
Sample Type
  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 Type: Whole Cells
    Application: Flow
  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 Type: Whole Cells
    Application: Flow
  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 Type: Whole Cells
    Application: Flow
  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 Type: Whole Cells
    Application: Flow

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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Isotype Controls
Description Application Cat# Citations Images  

Mouse IgG1 Fluorescein-conjugated Antibody

Ctrl IC002F 14  
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Staining Reagents
Description Application Cat# Citations Images  

Flow Cytometry Permeabilization/Wash Buffer I (1X)

Flow FC005 5
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Flow Cytometry Fixation Buffer (1X)

Flow FC004 3
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Flow Cytometry Fixation & Permeabilization Buffer Kit I

Flow FC009 3
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Flow Cytometry Fixation/Permeabilization Buffer I (1X)

Flow FC007 2
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