title: Profiling inflammatory responses with microfluidic immunoblotting.
abstract: Rapid profiling of signaling pathways has been a long sought after goal in biological sciences and clinical medicine. To understand these signaling pathways, their protein components must be profiled. The protein components of signaling pathways are typically profiled with protein immunoblotting. Protein immunoblotting is a powerful technique but has several limitations including the large sample requirements, high amounts of antibody, and limitations in assay throughput. To overcome some of these limitations, we have designed a microfluidic protein immunoblotting device to profile multiple signaling pathways simultaneously. We show the utility of this approach by profiling inflammatory signaling pathways (NFκB, JAK-STAT, and MAPK) in cell models and human samples. The microfluidic immunoblotting device can profile proteins and protein modifications with 5380-fold less antibody compared to traditional protein immunoblotting. Additionally, this microfluidic device interfaces with commonly available immunoblotting equipment, has the ability to multiplex, and is compatible with several protein detection methodologies. We anticipate that this microfluidic device will complement existing techniques and is well suited for life science applications.
authors: Michigan Nanotechnology Institute for Medicine and Biological Sciences, University of Michigan, Ann Arbor, Michigan, United States of America.
authors: Huai-Ning
authors: HN
authors: Chang
authors: Pascale R
authors: PR
authors: Leroueil
authors: Katherine
authors: K
authors: Selwa
authors: C J
authors: CJ
authors: Gasper
authors: Ryan E
authors: RE
authors: Tsuchida
authors: Jason J
authors: JJ
authors: Wang
authors: Walker M
authors: WM
authors: McHugh
authors: Timothy T
authors: TT
authors: Cornell
authors: James R
authors: JR
authors: Baker
authors: Sascha N
authors: SN
authors: Goonewardena
mesh_terms: D000818:Animals; D015153:Blotting, Western / Q000379:methods*; D002460:Cell Line; D006801:Humans; D007249:Inflammation / Q000378:metabolism*; D051379:Mice; D044085:Microfluidics / Q000379:methods*; D011506:Proteins / Q000378:metabolism*
chemical_list: D011506:Proteins