Rat TNF alpha ARExSet ELISA

Protocol Booklet

Key features and details

Species: Rat
Size: 5 plates,15 plates,50 plates
Sample Volume: 50 μl
Standard Curve Range: 31.25 pg/ml-2000 pg/ml
Assay type: Not Ready-to-Use
Sensitivity: 0.18 pg/ml(50 μl)
Operation Duration: 120min
Assay type: Sandwich Elisa

Brand:

CAT.NO. : ASER0007

US$ Please choose

Size:

5 plates 15 plates 50 plates

Trail, Bulk size or Custom requests Please contact us

Product Details

Background

Tumor necrosis factor alpha (TNF-alpha ), also known as cachectin and TNFSF1A, 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-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. 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). 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. 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. Shedding of membrane bound TNF-alpha by TACE/ADAM17 releases the bioactive cytokine, a 55 kDa soluble trimer containing the TNF-alpha extracellular domain. TNF-alpha binds the ubiquitous 55-60 kDa TNF RI and the hematopoietic cell-restricted 78-80 kDa TNF RII, both of which are also expressed as homotrimers. Both type I and type II receptors bind TNF-alpha with comparable affinity and can promote NFkB activation. Only TNF RI, however, contains a cytoplasmic death domain which triggers the activation of apoptosis. Soluble forms of both types of receptors are released into human serum and urine, and can neutralize the biological activity of TNF.

typical data

pg/mll	O.D.		Average	Corrected
0.00	0.0219	0.0214	0.0217
31.25	0.1501	0.1242	0.1372	0.1155
62.50	0.2717	0.2517	0.2617	0.2401
125.00	0.4979	0.4999	0.4989	0.4773
250.00	0.9518	0.9266	0.9392	0.9176
500.00	1.8560	1.7680	1.8120	1.7904
1000.00	3.1680	2.9560	3.0620	3.0404
2000.00	4.2224	4.1105	4.1665	4.1448

Product 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

Kit Content

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

Other Reagents Required

ARExSet^®Ancillary Reagent Kit (5 plates): containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and assay buffer.

96 well microplates: AREX Biosciences, Catalog # DSEP01.

Plate Sealers: AREX Biosciences, Catalog # DSSF01.

Coating Buffer: 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na₂HPO₄, 1.5 mM KH₂PO₄, pH 7.2-7.4, 0.2 μm filtered . AREX Biosciences, Catalog # DSCB01.

Blocking Buffer: AREX Biosciences, Catalog # DSBB01.

Wash Buffer: 0.05% Tween 20 in PBS5, pH 7.2-7.4. AREX Biosciences, Catalog # DSWB01.

Assay Buffer: 0.5%BSA, 0.05%Tween20, PBS Solution.AREX Biosciences, Catalog # DSAB01

Substrate Solution: Tetramethylbenzidine. AREX Biosciences, Catalog # DSTS01.

Stop Solution: 0.5 mol/ml H₂SO₄. AREX Biosciences, Catalog # DSSS01.

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