Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Primate - Papio cynocephalus (Yellow Baboon)

Applications

Validated:

Western Blot, Neutralization, Immunocytochemistry

Cited:

Immunohistochemistry, Western Blot, Neutralization, Immunocytochemistry, Affinity Assay, Bioassay, ELISA Development, Functional Assay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all TNF-alpha Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human TNF-alpha
Val77-Leu233
Accession # P01375

Specificity

Detects human TNF‑ alpha in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human TNF‑ alpha Antibody

Cytotoxicity Induced by TNF‑ alpha and Neutralization by Human TNF-alpha Antibody.

Cytotoxicity Induced by TNF‑ alpha and Neutralization by Human TNF-alpha Antibody.

Recombinant Human TNF-a (Catalog # 210-TA) induces cytotoxicity in the L-929 mouse fibroblast cell line in a dose-dependent manner (orange line). Cytotoxicity elicited by Recombinant Human TNF-a (1.5 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human TNF-a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-210-NA). The ND50 is typically 0.01-0.06 µg/mL.
TNF-a antibody in Human PBMCs by Immunocytochemistry (ICC).

TNF‑ alpha in Human PBMCs.

TNF-a was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Goat Anti-Human TNF-a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-210-NA) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Detection of Human TNF-alpha by Functional

Detection of Human TNF-alpha by Functional

TPM1 and TPM4 expression in response to TNF alpha.High molecular weight tropomyosin isoform expression in THP1 cells in response to elevated concentrations of TNF alpha. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27649540), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human TNF‑ alpha Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells

Western Blot

0.1 µg/mL
Sample: Recombinant Human TNF‑ alpha (Catalog # 210-TA)

Neutralization

Measured by its ability to neutralize TNF‑ alpha -induced cytotoxicity in the L‑929 mouse fibroblast cell line. Matthews, N. and M. L. Neale (1987) in Lymphokines and Interferons, A Practical Approach. Clemens, M. J. et al. (eds): IRL Press. 221. The Neutralization Dose (ND50) is typically 0.01-0.06 μg/ml in the presence of 1.5 ng/mL Recombinant Human TNF‑ alpha.

Reviewed Applications

Read 3 reviews rated 5 using AF-210-NA in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

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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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 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human TNF‑ alpha Antibody

For research use only

Citations for Human TNF‑ alpha Antibody

Customer Reviews for Human TNF‑ alpha Antibody (3)

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3 Customer Ratings
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Customer Images

Showing  1 - 3 of 3 reviews Showing All
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  • Human TNF-alpha Antibody
    Name: Stephanie Zandee
    Application: Immunohistochemistry
    Sample Tested: Adult brain
    Species: Human
    Verified Customer | Posted 04/13/2018
    Published in https://www.ncbi.nlm.nih.gov/pubmed/28169287 Used at 10ug/ml. Briefly, frozen brain sections were fixed in 4% PFA (Fisher Scientific), followed by antigen retrieval using heating in acid citric buffer (Vector, Burlingame, CA, USA). Endogenous avidin-biotin was blocked for 15 min (Vector). Sections were incubated with 10% horse serum in PBS (Biosera, Boussens, France) and Fc Receptor Blocking Solution was added (Human TruStain FcX Biolegend, London, UK). Primary antibody was added overnight at 4 °C and detected with donkey anti-goat-biotin (ab6578, Abcam), followed by streptavidin-alkaline phosphatase (SA-5100, Vector) and visualised with the Vector Blue Alkaline Phosphatase Substrate Kit III (Vector).
    Human TNF‑ alpha Antibody AF-210-NA
  • Human TNF-alpha Antibody
    Name: Balaji Mahender
    Application: ELISA
    Sample Tested: Heparin Plasma, EDTA Plasma and Citrate Plasma
    Species: Human
    Verified Customer | Posted 01/18/2018
  • Human TNF-alpha Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Serum and Plasma
    Species: Human
    Verified Customer | Posted 11/16/2017
    This antibody was used with MAB610 to build an ELISA to measure TNFa. This antibody was used as the detection while MAB610 was used as the capture.
    Human TNF‑ alpha Antibody AF-210-NA

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