The VCP inhibitor, CB-5083 (S8101), was from SelleckChem. The autophagy inhibitor, chloroquine diphosphate (CQ, 6628), the autophagosome-lysosome fusion inhibitor, and bafilomycin A1 (Baf-A1; Gibco, 19–148) were from Sigma-Aldrich. The following primary antibodies were used in this study: CHOP (2895), VCP (2648), LC3B (2775), and phospho-Histone H2A.X (Ser139, 9718) from Cell Signaling Technology. Lys48-specific ubiquitin antibody (05-1307) and vinculin (V9131) were from Sigma-Aldrich. The secondary antibodies, goat anti-rabbit IgG (31462) and goat anti-mouse IgG (31432), were from Invitrogen. pBABE-puro mCherry-EGFP-LC3B was a gift from Jayanta Debnath (Addgene plasmid #22418; http://n2t.net/addgene:22418 ; RRID: Addgene_22418) [45 (link)].
Goat anti mouse igg 31432
Manufactured by Thermo Fisher Scientific
Goat anti-mouse IgG (31432) is a secondary antibody conjugated with horseradish peroxidase (HRP) for the detection of mouse primary antibodies in various immunoassays. It is a purified goat-derived polyclonal antibody that binds to the Fc region of mouse immunoglobulin G (IgG).
Automatically generated - may contain errors
2 protocols using goat anti mouse igg 31432
1
VCP Inhibitor and Autophagy Modulators Protocol
2
Detecting PXDN in PFHR-9 and SMCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
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For the detection of PXDN in PFHR-9 cells, we developed a monoclonal antibody, adapting a previously described protocol [23 (link)]. A detailed description of the immunization and antibody production is provided in the supporting information.
For detection of PXDN in human SMCs we used a polyclonal PXDN specific antibody, previously developed in rabbit [17 (link)]. Primary antibodies used for western blots were purchased from Sigma Aldrich for detection of Flag-tag (mouse monoclonal Anti-FLAG M2 antibody, F3165), smooth muscle actin (mouse monoclonal antibody, A5228), and β-actin (mouse monoclonal antibody, A1978). HRP-conjugated secondary antibodies used for western blots, donkey anti-rabbit IgG (31458) and goat-anti-mouse IgG (31432), respectively, were purchased from Invitrogen.
For detection of PXDN in human SMCs we used a polyclonal PXDN specific antibody, previously developed in rabbit [17 (link)]. Primary antibodies used for western blots were purchased from Sigma Aldrich for detection of Flag-tag (mouse monoclonal Anti-FLAG M2 antibody, F3165), smooth muscle actin (mouse monoclonal antibody, A5228), and β-actin (mouse monoclonal antibody, A1978). HRP-conjugated secondary antibodies used for western blots, donkey anti-rabbit IgG (31458) and goat-anti-mouse IgG (31432), respectively, were purchased from Invitrogen.
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