Layered expression scanning: multiplex analysis of RNA and protein gels.
Northern blots and immunoblots are utilized in laboratories worldwide and offer several important features for analyzing mRNA and protein expression, including accuracy, low cost, evaluation of probe specificity, and information on transcript and protein forms based on molecular size. However, standard blotting techniques are hampered by three factors. They require a significant amount of input material, are laborious, and are capable of measuring only one protein or transcript at a time. Here we describe a simple yet effective technique for the multiplex analysis of standard RNA and protein gels using the layered expression scanning platform. The method relies on a novel membrane with high-affinity low-capacity binding characteristics. Using this approach, multiple blots from an RNA or protein electrophoresis gel can be simultaneously produced. We believe this method will be widely applicable to expression studies for a broad range of biological systems.
Tangrea
Michael A
MA
National Cancer Institute, Bethesda, MD, USA.
Flaig
Michael J
MJ
Ramesh
Arun
A
Best
Carolyn J M
CJ
Baibakov
Galina V
GV
Hewitt
Stephen M
SM
Mitchell
Capre D
CD
Hartmann
Dan-Paul
DP
Knezevic
Vladimir
V
Emmert-Buck
Michael R
MR
Chuaqui
Rodrigo F
RF
D015152:Blotting, Northern / Q000379:methods*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000295:instrumentation / Q000379:methods*; D015151:Immunoblotting / Q000379:methods*; D008567:Membranes, Artificial; D011506:Proteins / Q000032:analysis* / Q000378:metabolism*; D012313:RNA / Q000032:analysis* / Q000235:genetics*
D008567:Membranes, Artificial; D011506:Proteins; D012313:RNA