Cytoplasmic extract applications

This chapter describes and discusses methods used in our laboratory to investigate each step of male pronuclear formation in a sea urchin egg cytoplasmic extract. When available, procedures previously reported for surf clam male pro-nuclear assembly in vitro are also indicated. These protocols may be applicable to other marine invertebrates as well. 

In the domain of food industries, EED was used to reduce oxygen in fruit juice [46], to extract cytoplasmic proteins from alfalfa [47,48], to coagulate proteins [49], and to reduce disulfide bonds in proteins [50]. These applications are based on the electrode redox reactions coupled with monopolar membrane action. 

Retinoic acid receptors (RARs) have been detected m nuclear extracts from various cell types on the basis of their ability to bind radiolabeled retinoic acid (1 3). Prior to the isolation of RAR cDNAs in 1987 (4,5), biochemical evidence for the existence of nuclear retinoic-acid binding proteins distinct from cytoplasmic CRABP had been obtained (1). The techniques used in these experiments can be used to study the biochemical properties of the RARs and may also be applicable to the study of the related receptors known as RXRs, which bind 9-cis retinoic acid with high affinity (6). 

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