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10.1021/acs.analchem.6b00705 http://scihub22266oqcxt.onion/10.1021/acs.analchem.6b00705 C5113996!5113996!27270033     free     free     free Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Anal+Chem 2016 ; 88 (13): 6703-10 Nephropedia Template TP {{tp|p=27270033|t=2016. Multiplexed Western Blotting Using Microchip Electrophoresis |pdf=|usr=}}{{27270033}} gab.com Text Multiplexed Western Blotting Using Microchip Electrophoresis JO: Anal Chem http://www.kidney.de/pget.php?&tw=1&pmid=27270033 #FOAvip Western blotting is a commonly used protein assay that combines the selectivity of electrophoretic separation and immunoassay. The technique is limited by long time, manual operation with mediocre reproducibility, and large sample consumption, typically 10?20 ?g per assay. Western blots are also usually used to measure only one protein per assay with an additional housekeeping protein for normalization. Measurement of multiple proteins is possible; however, it requires stripping membranes of antibody and then reprobing with a second antibody. Miniaturized alternatives to Western blot based on microfluidic or capillary electrophoresis have been developed that enable higher-throughput, automation, and greater mass sensitivity. In one approach, proteins are separated by electrophoresis on a microchip that is dragged along a polyvinylidene fluoride membrane so that as proteins exit the chip they are captured on the membrane for immunoassay. In this work, we improve this method to allow multiplexed protein detection. Multiple injections made from the same sample can be deposited in separate tracks so that each is probed with a different antibody. To further enhance multiplexing capability, the electrophoresis channel dimensions were optimized for resolution while keeping separation and blotting times to less than 8 min. Using a 15 ?m deep × 50 ?m wide × 8.6 cm long channel it is possible to achieve baseline resolution of proteins that differ by 5% in molecular weight, e.g. ERK1 (44 kDa) from ERK2 (42 kDa). This resolution allows similar proteins detected by cross-reactive antibodies in a single track. We demonstrate detection of 11 proteins from 9 injections from a single Jurkat cell lysate sample consisting of 400 ng total protein using this procedure. Thus, multiplexed Western blots are possible without cumbersome stripping and reprobing steps. Twit Text FOAVip Multiplexed Western Blotting Using Microchip Electrophoresis ????Anal Chem http://www.kidney.de/pget.php?&tw=1&pmid=27270033 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=27270033 #FOAvip English Wikipedia <ref>{{Cite pmid|27270033}}</ref> Multiplexed Western Blotting Using Microchip Electrophoresis #MMPMID27270033 Jin S; Furtaw MD; Chen H; Lamb DT; Ferguson SA; Arvin NE; Dawod M; Kennedy RT Anal Chem 2016[Jul]; 88 (13): 6703-10 PMID27270033show ga Western blotting is a commonly used protein assay that combines the selectivity of electrophoretic separation and immunoassay. The technique is limited by long time, manual operation with mediocre reproducibility, and large sample consumption, typically 10?20 ?g per assay. Western blots are also usually used to measure only one protein per assay with an additional housekeeping protein for normalization. Measurement of multiple proteins is possible; however, it requires stripping membranes of antibody and then reprobing with a second antibody. Miniaturized alternatives to Western blot based on microfluidic or capillary electrophoresis have been developed that enable higher-throughput, automation, and greater mass sensitivity. In one approach, proteins are separated by electrophoresis on a microchip that is dragged along a polyvinylidene fluoride membrane so that as proteins exit the chip they are captured on the membrane for immunoassay. In this work, we improve this method to allow multiplexed protein detection. Multiple injections made from the same sample can be deposited in separate tracks so that each is probed with a different antibody. To further enhance multiplexing capability, the electrophoresis channel dimensions were optimized for resolution while keeping separation and blotting times to less than 8 min. Using a 15 ?m deep × 50 ?m wide × 8.6 cm long channel it is possible to achieve baseline resolution of proteins that differ by 5% in molecular weight, e.g. ERK1 (44 kDa) from ERK2 (42 kDa). This resolution allows similar proteins detected by cross-reactive antibodies in a single track. We demonstrate detection of 11 proteins from 9 injections from a single Jurkat cell lysate sample consisting of 400 ng total protein using this procedure. Thus, multiplexed Western blots are possible without cumbersome stripping and reprobing steps. äMultiplexed Western Blotting Using Microchip Electrophoresis (epmc).pdf DeepDyve Pubget Overpricing