Membrane and nuclear proteins were prepared from frozen livers as described (Engelking et al., 2004 (link); Moon et al., 2012 (link)). Equal aliquots (8 µg) of protein from individual livers were pooled (total, 40 µg) and the proteins were subjected to SDS-PAGE on 8% gels and transferred to nitrocellulose membrane (Bio-Rad, Hercules, CA). Immunoblot analyses were performed using polyclonal anti-mouse SREBP-1 and the monoclonal anti-mouse SREBP-2 antibody as described (Engelking et al., 2004 (link); McFarlane et al., 2015 (link); Moon et al., 2012 (link)) using rabbit monoclonal anti-SREBP-1 (IgG-20B12) and anti-SREBP-2 (IgG-22D5) antibodies that were generated against bacterially produced, His-tagged proteins containing amino acids 32–250 of mouse SREBP-1a or SREBP-2. Antibody-bound bands were detected using the SuperSignal West Pico Chemiluminescent Substrate system (ThermoScientific, cat. no. 34080). Anti-mouse CREB (cAMP response element binding protein, Invitrogen) and anti-dog Calnexin (Enzo Life Science, Farmingdale, NY) antibodies were used as loading controls for nuclear and membrane proteins, respectively.
Anti dog calnexin
Manufactured by Enzo Life Sciences
Sourced in United States
Anti-dog Calnexin is a primary antibody developed for the detection of calnexin, a protein involved in the folding and quality control of newly synthesized proteins in the endoplasmic reticulum of canine cells. This antibody can be used in various immunological techniques, such as Western blotting, immunoprecipitation, and immunohistochemistry, to study the expression and localization of calnexin in dog-derived samples.
Automatically generated - may contain errors
2 protocols using anti dog calnexin
1
Immunoblot Analysis of Liver Proteins
2
Protein Immunoblot Analysis of Mouse Liver
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Membrane and nuclear proteins were prepared individually from frozen livers, and equal amounts of protein from each mouse of the same group were pooled as previously described (Engelking et al., 2004 (link)). Aliquots of pooled proteins were subjected to SDS-PAGE on 8% gels and transferred to a nitrocellulose membrane (Bio-Rad, USA). Immunoblot analyses were performed using polyclonal anti-mouse SREBP-1, SREBP-2, and HMG-CoA reductase antibodies as previously described (Rong et al., 2017 (link)). Anti-Scavenger receptor class B type 1 (SR-BI) antibody was obtained from Abcam (USA) and anti-cluster of differentiation 36 (CD36) antibody was obtained from R&D Systems (USA). Anti-mouse cAMP response element binding protein (CREB; Invitrogen, USA) and anti-dog calnexin (Enzo Life Science, USA) antibodies were used as loading controls for nuclear and membrane proteins, respectively. Signals were detected using the SuperSignal West Pico Chemiluminescent Substrate System (Thermo Fisher Scientific) and visualized with Odyssey CLx Imaging system and LI-COR Image StudioTM software (LI-COR, USA).
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