Invitrogen™ Equine TNF alpha Uncoated ELISA Kit

Description

ELISA

The Equine TNF-alpha Uncoated ELISA Kit contains pre-matched antibody pairs and reagents for performing quantitative enzyme linked immunosorbent assays (ELISA) to detect and quantify protein levels of equine TNFalpha. Principle of the method ELISAs are designed to measure the amount of the target bound between a matched antibody pair. A target-specific antibody is coated to the bottom of the wells of a microplate, which is an overnight process. Samples, standards, or controls are then added into these wells and bind to the immobilized (capture) antibody. A sandwich is formed by the addition of the second (detector) antibody, a substrate solution is added that reacts with the enzyme-antibody-target complex to produce measurable signal. The intensity of this signal is directly proportional to the concentration of target present in the original specimen.

TNF alpha is a multifunctional proinflammatory cytokine that belongs to the tumor necrosis factor (TNF) superfamily. This cytokine is mainly secreted by macrophage and bind to its receptors, TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. TNF alpha is involved in the regulation of immune cells, cell proliferation, differentiation, apoptosis, lipid metabolism, and coagulation. TNF alpha exists as a multimer of two, three, or five noncovalently linked units, but shows a single 17 kDa band following SDS-PAGE under non-reducing conditions. Knockout studies in mice also suggested the neuroprotective function of TNF alpha, and has been observed to causes tumor necrosis when injected into tumor-bearing mice. Other functions of TNF-alpha include its role in the immune response to bacterial, viral, parasitic and certain fungal infections, as well as its role in the necrosis of specific tumors. TNF alpha causes cytolysis or cytostasis of certain transformed cells, being synergistic with interferon-gamma in its cytotoxicity. This cytokine has been implicated in a variety of diseases, including autoimmune diseases, insulin resistance, and cancer.

Specifications

• Accession Number: P29553
• Assay Range: 15.6 to 1000 pg/mL
• Assay Sensitivity: <15.6 pg/mL
• Description: Equine TNF alpha ELISA Reagent Kit
• Conjugate: HRP
• Immunoassay Kit Format: Colorimetric ELISA (TMB substrate) with Plates
• Product Type: ELISA
• Sample Type: Serum, Supernatant
• Specificity: No significant cross-reaction with bovine, mouse, or rat TNFa. Partial cross-reaction occurs with human and porcine TNFa (<10%)
• Sufficient For: 5 x 96 microplate wells
• Target Species: Equine
• For Use With (Equipment): Colorimetric Microplate Reader
• Gene ID (Entrez): 100033834
• Gene Symbol: TNF-a,TNF-alpha,TNFA,TNFSF2
• Kit Contents: Coating Antibody, Lyophilized Recombinant Standard, Detection Antibody, Streptavidin-HRP, Substrate Solution, Ready to use Stop Solution, 0.16M Sulfuric Acid, Certificate of Analysis: Lot-specific instructions for dilution of antibodies and standards
• Label or Dye: HRP
• Measurement: Absorbance at 450 nm minus absorbance (at 550 nm)
• Precision: Intra-assay CV <10%; inter assay CV <10%
• Quantity: 5 x 96 Tests
• Regulatory Status: RUO
• Sample Volume: Serum 100 μL, Supernatant 100 μL
• Storage Requirements: 2°C to 8°C
• Target: TNF alpha
• Test Time: 24 hrs
• Total Assay Time: 2 hrs 50 mins