Bcl-2 family proteins play critical roles in the regulation of programmed cell death and apoptosis (reviewed in refs 1-4). Changes in the levels or bioactivities of these proteins are associated with a variety of physiological processes where cell death occurs, including fetal development, haematopoietic and immune cell differentiation, oogenesis, mammary gland involution, and normal cell turnover in the epidermis, gut, and several other tissues. Moreover, pathological alterations in the expression of Bcl-2 family proteins have been documented in cancer, autoimmunity, immunodeficiency, heart failure, stroke, and other diseases (reviewed in refs 5-9). Consequently, methods for assessing the relative levels and bioactivities of Bcl-2 family proteins can be of interest to scientists in a board range of disciplines.
Here, we describe some of the methods routinely employed in our laboratory for this purpose. Many of the techniques described are specifically intended for analysis of the Bcl-2 (anti-apoptotic) or Bax (pro-apoptotic) proteins or mRNAs, but can be modified readily for applications to other members of the Bcl-2 family.
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