Immunohistochemistry: This antibody stained colchicine injected mouse brain (including the hippocampal fissure) tissue. The primary antibody was incubated at 1 mg/ml overnight at 4°C. This was followed by a peroxidase conjugated secondary antibody and then a fluorescein Tyramide Signal Amplification (TSA) reagent. Optimal concentrations and conditions may vary. Neutralization: To yield one-half maximal inhibition [ND] of the biological activity of mIL-1alpha (0.05 ng/ml), a concentration of 0 3-0.05 ug/ml of this antibody is required. ELISA: To detect mIL-1alpha 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-Murine IL-1alpha (LS-C104815
) as a detection antibody, allows the detection of at least 0.2-0.4 ng/well of recombinant mIL-1alpha. Western Blot: To detect mIL-1alpha by Western Blot analysis this antibody can be used at a concentration of 0.1-0.2 ug/ml. Used in conjunction with compatible secondary reagents the detection limit for recombinant mIL-1alpha is 1.5-3 ng/lane, under either reducing or non-reducing conditions.