• Assay Type

  Solid Phase Sandwich ELISA

• Format

  96-well strip plate

• Sample Type & Volume Required

  100 µL

• Range

  31.20 - 2,000 pg/mL

• Sufficient Materials

  For five or fifteen 96-well plates*

• Specificity

  Please see the

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

Ancillary Reagent Kit Available  

DY008, DuoSet ELISA Ancillary Reagent Kit 2 -

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human TNF-beta/TNFSF1. 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 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

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # ) 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 # ), 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 # ), or 0.05% Tween^® 20 in PBS, pH 7.2-7.4    

Reagent Diluent: (Catalog # ), 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 # )

Stop Solution: 2 N H₂SO₄ (Catalog # )

Microplates: R&D Systems (Catalog # )

Plate Sealers: ELISA Plate Sealers (Catalog # )

Preparation and Storage

• Storage

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

Background: Lymphotoxin-alpha/TNF-beta

Lymphotoxin alpha and Lymphotoxin beta are pro-inflammatory TNF superfamily ligands that play important roles in immune system development. LT-alpha can be secreted as a homotrimer that binds and activates TNF RI, TNF RII, HVEM, and TROY. It is required for development of the spleen, lymph nodes, and Peyer’s patches. It also regulates T cell homing to the gut and IgA induction. LT-alpha can additionally form membrane-associated heterotrimers with two copies of LT-beta/TNF-beta on the surface of B, T, LTi, and ILC3 cells. This heterotrimer binds and activates the Lymphotoxin beta R on macrophages, dendritic cells, hepatocytes, intestinal epithelial cells (IEC), follicular dendritic cells (FDC), and high endothelial venules (HEV). The heterotrimer promotes the development of FDC networks and HEV in lymphoid tissue, the class switching of immature B cells for IgA production, and the production of homeostatic IL-22 by IEC.

• Entrez Gene IDs:

  4049 (Human); 16992 (Mouse);

• Long Name:

  Tumor Necrosis Factor beta

• Aliases:

  LT; LTA; LT-alpha; lymphotoxin alpha (TNF superfamily, member 1); Lymphotoxin alpha; Lymphotoxinalpha; Lymphotoxin-alpha; tnfb; TNF-beta; TNFBlymphotoxin-alpha; TNFSF1; TNFSF1B; TNFSF1TNF-beta; tumor necrosis factor beta; Tumor necrosis factor ligand superfamily member 1