Protocol for cytostatic factor extract preparation for spindle assembly. The quality of the eggs is essential for good CSF extracts. Always sacrifice quantity for quality when trying to make functional extracts. Discard any batches of eggs that have "puff balls" or activated eggs that constitute more than 10% of the eggs. Use laid eggs and collect eggs at about 16 to 17 hours after priming with Progesterone (found in Pregnant Mare Serum Gonadotropin (PMSG)). - [Read Cytostatic Factor (CSF) Extract Preparation for Xenopus Spindle Assembly Protocol]
Protocol describes a method for removal of mouse ovaries in combination with the administration of progesterone, which causes the arrest of blastocyst development. This procedure is performed during the afternoon of the third day of pregnancy to ensure that the morulae have moved sufficiently into the oviducts and will be less likely to be damaged during the ovariectomy. - [Read Ovariectomy for Induction of Blastocyst Implantation Delay Protocol]
In Vitro Translated Xenopus Mos Kinase Assay Protocol. In response to progesterone, immature Xenopus oocytes mature to eggs that can be fertilized. The Mos protein kinase is essential for oocyte maturation, most likely due to its ability to activate the MAP kinase cascade. This MAP kinase cascade eventually leads to the activation of Cdc2/cyclin B and entry into M phase. In this protocol, tagged Mos kinase is translated in vitro, immunopurified, and used in a kinase assay.