[Evaluation of binding of FITC-prolactin conjugate to prolactin receptor]

ABSTRACT

Prolactin is an important mammalian hormone and the associated receptor is recognized in many different cells. Radioligand and histochemical methods are both used for assaying prolactin receptor. To produce FITC-prolactin conjugate and also to study the ability of conjugate to bind prolactin receptor. In this experimental study FITC was bound to prolactin in alkaline solution. FITC-prolactin conjugate was separated from free FITC by chromatographic method. Later, the ability of FITC-prolactin conjugate to bind prolactin receptor of peripheral blood mononuclear cells [PBMC] was assessed by flowcytometry. Fluorescence emission was detected in 2.1% of the cells in absence of FITC-prolactin. Following the addition of FITC-prolactin conjugate to cells for one hour and further washing, the fluorescence emission was detected in 27.8% of cells. For PBMC, these data were 0.02% and 11.8%, respectively. Regarding the data obtained in our study, FITC-prolactin conjugate can bind prolactin receptor. Therefore, this conjugate could be used for assessing prolactin receptor by fluorometric method: