Typically, 10% resolving gel of SDS-PAGE was performed unless otherwise indicated. One of the resulting gels was stained with Coomassie colloid blue 44 (link), and the other gel was subjected to electrophoretic transfer to hybond-C membrane and immunoblotting 50 (link). Signals on the hybond-C membrane were visualized with an enhanced chemiluminescence kit. Nongradient blue native gel electrophoresis was performed as previously described 41 (link). All images were scanned by an EPSON PERFECTION 1670 scanner. All densitometric quantifications of gel images were also analyzed by AlphaEaseFC software.
Perfection 1670 scanner
Manufactured by Epson
The Epson PERFECTION 1670 is a flatbed scanner designed for laboratory use. It features a 1600 x 3200 dpi optical resolution and can scan documents up to 8.5 x 11.7 inches in size. The scanner is capable of 48-bit color depth and supports various file formats.
Automatically generated - may contain errors
5 protocols using perfection 1670 scanner
1
SDS-PAGE, Immunoblotting, and Blue Native Gel Analysis
2
Protein Characterization via Electrophoresis
3
Protein Purification and Analysis
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SDS-PAGE (typically 10% resolving gel) was performed according to standard procedures [43] (link). One of the resulting gels was stained with Coomassie colloid blue [38] (link), and the other gel was subjected to electrophoretic transfer to membrane for immunoblotting [44] (link). Signals on the immunoblotting membrane were visualized with an enhanced chemiluminescence kit. Nongradient blue native gel electrophoresis (BN-PAGE) was performed as previously described [36] (link). All images were scanned by an EPSON PERFECTION 1670 scanner. All densitometric quantifications of gel images were analyzed by AlphaEaseFC software.
4
Multiplex Imaging and Quantification
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Blots from Figs 1 –6 were obtained by autoradiography using GE Healthcare Amersham HyperfilmTM ECL. Western blots were scanned using Epson Perfection 1670 Scanner and quantified using an image processor software (ImageJ). Blots from Fig. 7 and Supplementary Figs 4–5 were obtained using the Odyssey Infrared Imaging System (LI-COR Biosciences). BRET experiments were performed using the Mithras LB 940 (Berthold technologies). Uptiblue measurements were performed using a TyphoonTM FLA 9000 (GE Healthcare).
5
Protein Detection and Imaging Protocol
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Blots from Figs. 1 , 3 and Supplementary Figs. 2 –4 were obtained by autoradiography using GE Healthcare Amersham HyperfilmTM ECL. Western blots were scanned using Epson Perfection 1670 Scanner. In vivo imaging (Figs. 1a and 2e ) was performed using Biospace Photon Imager and analyzed with Biospace software (Biospace, Nesles-la-Vallée, France).
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