A Novel Immunological Assay for Hepcidin Quantification in Human Serum
Figure 1
Specificity of the polyclonal antibody against native hepcidin.
(A) Immunohistochemical staining of liver tissue sections using the polyclonal antibody against hepcidin25-His (a-Hep25). Secondary anti-rabbit antibody was used as negative control (negative). The specificity of the polyclonal antibody was verified after reduction of the signal following preincubation with hepcidin25-His. (B) Western blot analysis of serum proteins less than 30 kDa. 10 ml of serum was filtered through a 30 kDa filter, and the filtrate was precipitated with 25% TCA. Precipitated proteins were subjected to electrophoresis on a 4–12% Nu-PAGE gel, followed by Western blot using the polyclonal antibody against hepcidin25-His.