Caspases,Paracaspases, and Metacaspases Methods and Protocols

108 References Sulston JE, Horvitz HR (1977) Post- embryonic cell lineages of the nematode, Caenorhabditis elegans. Dev Biol 56 ...

109 Peter V. Bozhkov and Guy Salvesen (eds.), Caspases, Paracaspases, and Metacaspases: Methods and Protocols, Methods in Molecu ...

110 traditional cell death and include a plethora of non-apoptotic func- tions. Caspases have been implicated in cell migration ...

111 2 Materials Many fl y stocks can be obtained from the Bloomington Drosophila Stock Center ( http://fl ystocks.bio.indiana.e ...

112 3 Methods This dissection technique is described assuming right hand domi- nance. Each person will likely fi nd a variation ...

113 Using #55 forceps carefully transfer samples to a glass slide with a drop of VECTASHIELD ( see Note 14 ). Using the dissect ...

114 If the parental cross was set up such that dominant larval markers such as Tubby or a GFP reporter will positively or neg- ...

115 Take care to make sure that each antibody was raised in a different species which will prevent cross-reaction during the sec ...

116 Kumar S (2007) Caspase function in pro- grammed cell death. Cell Death Differ 14(1):32–43. doi: 10.1038/sj.cdd.4402060 Xu D ...

117 Fan Y, Bergmann A (2010) The cleaved-Caspase- 3 antibody is a marker of Caspase-9-like DRONC activity in Drosophila. Cell D ...

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119 Peter V. Bozhkov and Guy Salvesen (eds.), Caspases, Paracaspases, and Metacaspases: Methods and Protocols, Methods in Molecu ...

120 and discovering that the Drosophila melanogaster protein, reaper, could function in a vertebrate system to induce apoptosis ...

121 Exogenous sperm nuclei can even be added to X. laevis egg extract, and nuclear morphology can be studied as a technique for ...

122 X. laevis egg extract. 2.5-mL (11 × 34 mm) polycarbonate centrifuge tubes (e.g., Beckman). TLS ultracentrifuge and TLS-55 s ...

123 X. laevis egg extract. 20× EM. Centrifuge fi lter, PVDF 0.1 μm (Millipore, cat. # UFC30VVNB or similar). SDS sample buffer. ...

124 Mature female primed frogs. 10 % benzocaine. Dissection equipment. Liberase blendzyme 3 (Roche, Switzerland) or similar. OR ...

125 extract and whole oocytes, and how to analyze targets of execu- tioner caspase activation including DNA fragmentation and po ...

126 Wash eggs three times in 1× ELB. At this stage, bad or poor quality eggs can be removed. Mature, good quality, X. laevis eg ...

127 Transfer the crude egg extract prepared as described in the previous section to a 2.5-mL polycarbonate centrifuge tube. All ...