Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Flow Cytometry

Cited:

Flow Cytometry

Label

Fluorescein (Excitation = 488 nm, Emission = 515-545 nm)

Antibody Source

Monoclonal Mouse IgG1 Clone # 6401
View all TNF-alpha Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human TNF-alpha
Gly57-Leu233 (predicted)
Accession # P01375

Specificity

Detects cell surface expressed TNF-alpha (membrane and receptor bound forms) by flow cytometry. Detects human TNF-alpha in direct ELISAs and Western blots. In direct ELISAs, approximately 25‑50% cross‑reactivity with recombinant porcine TNF‑ alpha and recombinant rhesus macaque TNF-alpha is observed but no cross-reactivity with recombinant cotton rat TNF‑ alpha, recombinant rat TNF‑ alpha, recombinant human (rh) LT alpha 1/ beta 2, rhLT alpha 2/ beta 1, rhAPRIL, rhBAFF, rhEDA‑A2, recombinant mouse EDA, rhFas Ligand, rhLIGHT, rhOX40 Ligand, rhTRAIL, rhTRANCE, rhTWEAK, or rhVEGI is observed.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Human TNF‑ alpha Membrane Form Fluorescein‑conjugated Antibody

Detection of TNF‑ alpha in PBMC monocytes by Flow Cytometry.

Stimulated PBMC monocytes treated with 1 μg/mL LPS at 37 degrees Celsius for 4 hours were stained with Mouse Anti-Human CD14 APC‑conjugated Monoclonal Antibody (Catalog # FAB3832A) and either (A) Mouse Anti-Human TNF‑ alpha Membrane Form Fluorescein‑conjugated Monoclonal Antibody (Catalog # FAB210F) or (B) Mouse IgG1 Fluorescein Isotype Control (Catalog # IC002F). View our protocol for Staining Membrane-associated Proteins.

Applications for Human TNF‑ alpha Membrane Form Fluorescein‑conjugated Antibody

Application
Recommended Usage

Flow Cytometry

10 µL/106 cells
Sample: Human peripheral blood monocytes and stimulated PBMC monocytes treated with 100 ng/mL LPS at 37 degrees Celsius for 4 hours

Reviewed Applications

Read 1 review rated 5 using FAB210F in the following applications:

Spectra Viewer

Plan Your Experiments

Use our spectra viewer to interactively plan your experiments, assessing multiplexing options. View the excitation and emission spectra for our fluorescent dye range and other commonly used dyes.

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from ascites

Formulation

Supplied in a saline solution containing BSA and Sodium Azide.

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

Alternate Names

Cachetin, DIF, TNF, TNF-A, TNFA, TNFalpha, TNFG1F, TNFSF1A, TNFSF2

Entrez Gene IDs

7124 (Human); 21926 (Mouse); 24835 (Rat); 397086 (Porcine); 280943 (Bovine); 403922 (Canine); 102139631 (Cynomolgus Monkey); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit)

Gene Symbol

TNF

UniProt

P01375

Additional TNF-alpha Products

Product Documents for Human TNF‑ alpha Membrane Form Fluorescein‑conjugated Antibody

Certificate of Analysis

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Product Specific Notices for Human TNF‑ alpha Membrane Form Fluorescein‑conjugated Antibody

For research use only

Citations for Human TNF‑ alpha Membrane Form Fluorescein‑conjugated Antibody

Customer Reviews for Human TNF‑ alpha Membrane Form Fluorescein‑conjugated Antibody (1)

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  • Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: See PMID: 21109521
    Species: Human
    Verified Customer | Posted 01/22/2015

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FAQs for Human TNF‑ alpha Membrane Form Fluorescein‑conjugated Antibody

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  • Q: Why do FAB210 and IC210 give different intracellular staining results?

    A: 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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