Does the Mouse TNF-alpha DuoSet ELISA recognize the monomer and/or trimer form of TNF-alpha?

Name: Mouse TNF-alpha DuoSet ELISA
Brand: R&D Systems
SKU: DY410
Rating: 5 (24 reviews)

TNF-alpha self-dissociates to monomers in the picomolar range. The assay standard curve concentration is from approximately 2 - 118 femtomolar, the range in which self-dissociation is expected to occur. This supports that the assay detects monomeric TNF-alpha.

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R&D Systems, part of Bio-Techne | Catalog # DY410

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Product Summary
Product Specifications
Scientific Data
Kit Contents
Other Reagents Required
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Assay Type

Solid Phase Sandwich ELISA

Assay Range

31.2-2000 pg/mL

Sample Type

Cell culture supernates, serum, and plasma
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet

Reactivity

Mouse

Mouse TNF-alpha DuoSet ELISA Features

Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
Development protocols are provided to guide further assay optimization
Assay can be customized to your specific needs
Economical alternative to complete kits

View all TNF-alpha ELISA Kits »

Product Summary for Mouse TNF-alpha DuoSet ELISA

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant mouse TNF-alpha. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Specifications

Assay Format

96-well strip plate (sold separately)

Sample Volume Required

100 µL

Detection Method

Colorimetric ELISA - 450nm (TMB)

Conjugate

Biotin

Specificity

Please see the product datasheet

Label

HRP

Scientific Data Images for Mouse TNF-alpha DuoSet ELISA

Mouse TNF-alpha ELISA Standard Curve

Kit Contents for Mouse TNF-alpha DuoSet ELISA

Capture Antibody
Detection Antibody
Recombinant Standard
Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na₂HPO₄, 1.5 mM KH₂PO₄, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween^® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H₂O₂) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H₂SO₄ (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Preparation and Storage

Shipping

The product is shipped at ambient temperature. 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.

Background: TNF-alpha

Tumor necrosis factor alpha (TNF-alpha), also known as cachectin and TNFSF1A, is the prototypic ligand of the TNF superfamily (1). It is a pleiotropic molecule that plays a central role in inflammation, immune system development, apoptosis, and lipid metabolism (2-5). TNF-alpha is also involved in a number of pathological conditions including asthma, Crohn's disease, rheumatoid arthritis, neuropathic pain, obesity, type 2 diabetes, septic shock, autoimmunity, and cancer (5-11).

Mouse TNF-alpha is synthesized as a 26 kDa type II transmembrane protein that consists of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 179 aa extracellular domain (ECD) (12). Within the ECD, mouse TNF-alpha shares 95% aa identity with rat, and 80% aa identity with canine, equine, feline, human, rabbit, and porcine TNF-alpha. It 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 (13). Cell surface TNF-alpha can both induce the lysis of tumor cells and virus infected cells, and generate its own downstream cell signaling following ligation by soluble TNF RI (14, 15). Shedding of membrane bound TNF-alpha by TACE/ADAM17 releases the bioactive cytokine, a 55 kDa soluble trimer containing the TNF-alpha extracellular domain (16-18).

TNF-alpha binds the ubiquitous 55-60 kDa TNF RI (19, 20) and the hematopoietic cell-restricted 78-80 kDa TNF RII (21, 22), both of which are also expressed as homotrimers (1, 23). Both type I and type II receptors bind TNF-alpha with comparable affinity and can promote NFkB activation (24-27). Only TNF RI, however, contains a cytoplasmic death domain which triggers the activation of apoptosis (3, 28). Soluble forms of both types of receptors are released into human serum and urine, and can neutralize the biological activity of TNF (29-31).

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

Additional TNF-alpha Products

Product Documents for Mouse TNF-alpha DuoSet ELISA

Product Datasheets

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COA

SDS

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Product Specific Notices for Mouse TNF-alpha DuoSet ELISA

For research use only

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