Abstract
Publisher Summary This chapter discusses the measurement of hormone secretion from individual cells by cell blot assay. In the cell blot assay, cells are incubated on, and bound to, a transfer membrane. Hormone released from the cells is captured on the membrane. The membrane is then processed according to Western blot methodology. This includes incubation in primary antiserum, secondary antiserum to which an enzyme is conjugated, and enzyme substrate. The released hormone product surrounding the hormone-secreting cells (e.g., prolactin and growth hormone) becomes visible as a zone of secretion immediately encompassing the cell. This secretion zone is then analyzed and quantified. Heterogeneity of hormone release per cell becomes apparent. One of the limitations of the cell blot assay is the inability to visualize the nonsecreting cells on the immobilon membrane. Advantages of the cell blot assay include its simplicity and rapidity, use of dilute preparations of antisera, objectivity in measurement of hormone secretion, and capability of quantification of hormone release from single cells.
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