Caspases,Paracaspases, and Metacaspases Methods and Protocols

248 Metacaspase activity in vivo can be also assessed by analyzing the effi ciency of natural substrate cleavage, e.g., cleavage ...

249 Rinse the membrane in PBST buffer and incubate with primary antibody for 1 h at room temperature or overnight at 4 °C. Wash ...

250 Produce the protein substrate in the cell free TNT ® Coupled Transcription/Translation System (TNT ® mix) according the man ...

251 4 Notes P1 refers to the residue situated N-terminus to the scissile bond cleaved by the protease extending towards the N-t ...

252 Uren AG, O’Rourke K, Aravind L et al (2000) Identifi cation of paracaspases and metacas- pases: two ancient families of cas ...

253 Arabidopsis metacaspase 2d. J Biol Chem 286: 10027–10040 Tsiatsiani L, Timmerman E, De Bock P-J et al (2013) The Arabidopsi ...

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

256 DIagonal Chromatography (COFRADIC, [ 5 ]). More recently, the Overall lab and our lab have published positional proteomics a ...

257 α-amino group (in vivo) or a butyrylated α-amino group (in vitro), and non N-terminal peptides (i.e., internal peptides and ...

258 Tris(2-carboxyethyl)phosphine (TCEP) stock solution: 150 mM TCEP ( see Note 1 ). Iodoacetamide stock solution: 300 mM iodoa ...

259 Repeat steps 4 and 5 ( see Note 6 ). Add 1.25 μL of 1 M glycine to quench the non-reacted N -hydroxysuccinimide esters. Inc ...

260 Plasman K et al (2013) Contemporary posi- tional proteomics strategies to study protein processing. Curr Opin Chem Biol 17: ...

261 Staes A et al (2008) Improved recovery of proteome- informative, protein N-terminal peptides by combined fractional diagona ...

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

CED-9 ............................................................. 102, 105, 106 Cell free system ............................. ...

LmjMCA ................................................. 1 90–193, 196–200 Luminophore ......................................... ...

Stokes’ shift ........................................................................3 1 Strong cation exchange chromatography ...