Immunohistochemistry: This antibody stained formalin-fixed, paraffin-embedded sections of human breast invasive ductal carcinoma. The recommended concentrations are 2.5 ug/ml-5 ug/ml with a two-hour incubation at room temperature or an overnight incubation at 4°C. An HRP-labeled polymer detection system was used with a DAB chromogen. This antibody was also tested in an HRP-biotin labeling detection system. Heat induced antigen retrieval with a pH 6 Sodium Citrate buffer is recommended. Optimal concentrations and conditions may vary. Neutralization: To yield one-half maximal inhibition [ND] of the biological activity of Human TNF-alpha (0.25 ng/ml), a concentration of 0.012-0.014 ug/ml of this antibody is required. ELISA: To detect Human TNF-alpha by sandwich ELISA (using 100 ul/well antibody solution) a concentration of 0.5-2 ug/ml of this antibody is required. This antigen affinity purified antibody, in conjunction with Biotinylated Anti-Human TNF-alpha (
LS-C104788) as a detection antibody, allows the detection of at least 0.2-0.4 ng/well of recombinant Human TNF-alpha. Western Blot: To detect Human TNF-alpha by Western Blot analysis this antibody can be used at a concentration of 0.1-0.2 ug/ml. When used in conjunction with compatible secondary reagents, the detection limit for recombinant Human TNF-alpha is 1.5-3 ng/lane, under either reducing or non-reducing conditions.