Esophageal Adenocarcinoma Methods and Protocols

277 Apply the chemiluminescent substrate to the membrane and incubate at dark for 2–5 min. Capture the chemiluminescent signals ...

278 For stable clone selection with puromycin, sufficient amount of antibiotic should be used to kill all the non-transfected c ...

279 Hong J, Resnick M, Behar J et al (2011) Role of Rac1 in regulation of NOX5-S function in Barrett’s esophageal adenocarcinom ...

281 Alfred K. Lam (ed.), Esophageal Adenocarcinoma: Methods and Protocols, Methods in Molecular Biology, vol. 1756, https://doi. ...

282 fine capillary, producing a fine aerosol of charged particles in an electric field, or matrix-assisted laser desorption ioni ...

283 2 Materials U9 Buffer: 9 M urea, 2% CHAPS, 3[(3-cholamidopropyl) dimethylammonio)]-propanesulfonic acid. Prepare equilibrat ...

284 ●● IMAC—immobilized metal affinity capture. These can be activated using copper or nickel. ●● CM10—weak cation exchanger. Th ...

285 ples, tumor tissue lysates, and fractionated samples prepared for protein identification purposes. The technique involves: ...

286 further washes with equilibration buffer without triton (2 × 150 μL buffer and 5 min incubations). Perform a final wash wit ...

287 Wash each spot twice with 5 μL of water using the on/off pipette technique (see Note 4). Allow the spots to dry. Add sinapi ...

288 For quantitative analysis, collate spectra obtained for each sam- ple group being tested into (high or low molecular weight ...

289 This technique is as an alternative to anion exchange chromatogra- phy. Isoelectric focusing separates proteins according to ...

290 4 Notes Do not use serum and plasma interchangeably. The clotting process will remove proteins from serum. Hence, a decisio ...

291 using, e.g., a 20% peak threshold and a signal/noise ratio of 3. Once an appropriate peak threshold and signal/noise ratio i ...

292 now become available for this application [ 11 ]. More recently, concern has been expressed that removal of carrier proteins ...

293 References Cahill DJ (2001) Protein and antibody arrays and their medical applications. J Immunol Methods 250:81–91 Plebani ...

295 Alfred K. Lam (ed.), Esophageal Adenocarcinoma: Methods and Protocols, Methods in Molecular Biology, vol. 1756, https://doi. ...

296 EsophagEal adEnocarcinoma: mEthods and protocols Index S Sanger sequencing ����������������������217, 219, 220, 222, 223, 22 ...