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Human TNF-alpha PE-conjugated Antibody

Catalog #: IC210P
9 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
IC210P
Detection of TNF‑ alpha in Human PBMCs by Flow Cytometry.
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Citations (9)
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Human TNF-alpha PE-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
Phycoerythrin (Excitation= 488 nm, Emission= 565-605 nm)

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.

Scientific Data

Intracellular Staining by Flow Cytometry Detection of TNF- alpha antibody in Human PBMCs antibody 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 were stained with (A) Mouse Anti-Human TNF-alpha PE-conjugated Monoclonal Antibody (Catalog # IC210P) or (B) isotype control antibody (IC002P) 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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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
Store the unopened product at 2 - 8° C. Do not use past expiration date. Protect from light.

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); 102139631 (Cynomolgus Monkey); 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

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Citations for Human TNF-alpha PE-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.

9 Citations: Showing 1 - 9
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  1. Serum microRNA181a: Correlates with the intracellular cytokine levels and a potential biomarker for acute graft-versus-host disease
    Authors: Linna Xie
    Cytokine, 2016-06-08;85(0):37-44.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  2. Expansion of anti-mesothelin specific CD4+ and CD8+ T cell responses in patients with pancreatic carcinoma.
    Authors: Chen Y, Ayaru L, Mathew S, Morris E, Pereira S, Behboudi S
    PLoS ONE, 2014-02-10;9(2):e88133.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  3. NK cells from HCV-infected patients effectively induce apoptosis of activated primary human hepatic stellate cells in a TRAIL-, FasL- and NKG2D-dependent manner.
    Authors: Glassner A, Eisenhardt M, Kramer B, Korner C, Coenen M, Sauerbruch T, Spengler U, Nattermann J
    Lab. Invest., 2012-03-26;92(7):967-77.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  4. Acetaldehyde stimulates monocyte adhesion in a P-selectin- and TNFalpha-dependent manner.
    Authors: Redmond EM, Morrow D, Kundimi S, Miller-Graziano CL, Cullen JP
    Atherosclerosis, 2008-10-18;204(2):372-80.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  5. Human CD4(+) T cells recognize an epitope within alpha-fetoprotein sequence and develop into TGF-beta-producing CD4(+) T cells.
    Authors: Alisa A, Boswell S, Pathan AA, Ayaru L, Williams R, Behboudi S
    J. Immunol., 2008-04-01;180(7):5109-17.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  6. Interaction between transmembrane TNF and TNFR1/2 mediates the activation of monocytes by contact with T cells.
    Authors: Rossol M, Meusch U, Pierer M, Kaltenhauser S, Hantzschel H, Hauschildt S, Wagner U
    J. Immunol., 2007-09-15;179(6):4239-48.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  7. Analysis of CD4+ T-Cell responses to a novel alpha-fetoprotein-derived epitope in hepatocellular carcinoma patients.
    Authors: Alisa A, Ives A, Pathan AA, Navarrete CV, Williams R, Bertoletti A, Behboudi S
    Clin. Cancer Res., 2005-09-15;11(18):6686-94.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  8. Different domains of Pseudomonas aeruginosa exoenzyme S activate distinct TLRs.
    Authors: Epelman S, Stack D, Bell C, Wong E, Neely GG, Krutzik S, Miyake K, Kubes P, Zbytnuik LD, Ma LL, Xie X, Woods DE, Mody CH
    J. Immunol., 2004-08-01;173(3):2031-40.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  9. Lack of T Cell Response to iPSC-Derived Retinal Pigment Epithelial Cells from HLA Homozygous Donors
    Authors: Sunao Sugita, Yuko Iwasaki, Kenichi Makabe, Takafumi Kimura, Takaomi Futagami, Shinji Suegami et al.
    Stem Cell Reports

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.

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