Mouse TNF-Alpha ELISA Assay

Product Overview:

The mouse TNF-alpha ELISA assay is a highly sensitive two-site enzyme linked immunoassay (ELISA) for measuring TNF-a in biological samples of mice.

Assay Principle:

In this mouse TNF-alpha ELISA assay the TNF-a present in samples reacts with the anti-TNF-a antibodies which have been adsorbed to the surface of polystyrene microtitre wells. After the removal of unbound proteins by washing, the Detection Antibody, biotin conjugated anti-TNF-a is added and complexes are formed. Following a wash step, the horseradish peroxidase (HRP) conjugated Streptavidin is added and complexes are formed. After another washing step, the complexes are assayed by the addition of a chromogenic substrate, 3,3’,5,5’-tetramethylbenzidine (TMB). The quantity of bound enzyme varies directly with the concentration of TNF-a in the sample tested; thus, the absorbance, at 450 nm, is a measure of the concentration of TNF-a. The quantity of TNF-a in the mouse sample can be interpolated from the standard curve constructed from the standards, and corrected for sample dilution.

Dynamic Range:

31.2 pg/ml – 2000 pg/ml

Incubation Time:

3 hours 10 minutes

Sample Type:

Cell culture supernates, urine, serum and plasma

Sample Size:

100 µl

Specimen Collection and Handling:

Blood should be collected by venipuncture. The serum should be separated from the cells after clot formation by centrifugation. For plasma samples, blood should be collected into a container with an anticoagulant and then centrifuged. Care should be taken to minimize hemolysis, excessive hemolysis can impact your results. Assay immediately or aliquot and store samples at -20°C. Avoid repeated freeze-thaw cycles.