Immunohistochemistry: This antibody stained formalin-fixed, paraffin-embedded sections of human colon/rectum adenocarcinoma. The recommended concentration is 0.5 ug/ml with an overnight incubation at 4°C. An HRP-labeled polymer detection system was used with a DAB chromogen. Heat induced antigen retrieval with a pH 6 sodium citrate buffer is recommended. Optimal concentrations and conditions may vary. Tissue samples were provided by the Cooperative Human Tissue Network, which is funded by the National Cancer Institute. Neutralization: To yield one-half maximal inhibition [ND] of the biological activity of hMIP-5 (50 ng/ml), a concentration of 1.75-3 ug/ml of this antibody is required. ELISA: To detect hMIP-5 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 MIP-5 (
LS-C104743) as a detection antibody, allows the detection of at least 0.2-0.4 ng/well of recombinant hMIP-5. Western Blot: To detect hMIP-5 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 hMIP-5 is 1.5-3 ng/lane, under either reducing or non-reducing conditions.