Constitution and quantity of lysis buffer alters outcome of reverse phase protein microarrays.
Application of novel technology to clinical samples requires optimization of procedures. Reverse phase protein lysate arrays use femtomolar quantities of tissue lysate from clinical samples with which to profile biochemical events happening in the tumor. We analyzed the effects of different tissue solubilization buffers on frozen ovarian tumor samples in order to identify the system with the best signal intensity dynamic range, reproducibility, tissue solubility, and signal consistency. A modified RIPA-like buffer supplemented with DTT and SDS was deemed optimal.
Winters
Mary
M
Laboratory of Pathology, Center for Cancer Research, National Cancer Institute, Bethesda, MD, USA.
Dabir
Bhavana
B
Yu
Minshu
M
Kohn
Elise C
EC
D002021:Buffers; D004229:Dithiothreitol / Q000737:chemistry*; D005260:Female; D006801:Humans; D009363:Neoplasm Proteins / Q000032:analysis*; D010051:Ovarian Neoplasms / Q000175:diagnosis*; D040081:Protein Array Analysis / Q000379:methods*; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry*; D012995:Solubility
D002021:Buffers; D009363:Neoplasm Proteins; D012967:Sodium Dodecyl Sulfate; D004229:Dithiothreitol