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Horseradish peroxide hrp conjugated anti mouse igg

Manufactured by Santa Cruz Biotechnology

Horseradish peroxide (HRP)-conjugated anti-mouse IgG is a secondary antibody used in immunoassays and other techniques to detect the presence of mouse immunoglobulin G (IgG) in samples. The HRP enzyme label allows for colorimetric or chemiluminescent detection of the bound antibody.

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2 protocols using horseradish peroxide hrp conjugated anti mouse igg

1

Quantification of ENO1 Protein Levels

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PDAC cells (1×107) from the various cell lines were harvested, lysed, resolved and transferred to nitrocellulose membranes, as previously described [40 (link)]. Membranes were incubated for 1 h at RT with anti-ENO1 72/1 mAb or rabbit polyclonal anti- β-Actin antibody (Sigma-Aldrich), at dilutions of 1:2000 in Tween-Tris-Buffered Saline (TTBS) and then probed with a horseradish peroxide (HRP)-conjugated anti-mouse IgG (Santa Cruz) or HRP-conjugated goat anti-rabbit Ig secondary antibody (Sigma-Aldrich) at dilutions of 1:2000. For Western blot analysis of ENO1 in PANC-1/P, PANC-1/M and HPDE cells, membranes were probed with in-house purified rabbit antiserum against ENO1 or with mouse antibody specific to β-Actin (Sigma St. Louis, MO, USA) as a protein loading control. Immunocomplexes were detected by probing with appropriate secondary antibodies conjugated with HRP (Jackson ImmunoResearch), and were visualized using the SuperSignal detection system (Thermo Fisher, Waltham, MA, USA).
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2

Western Blot Analysis of BAF Subunits

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Lysates were prepared using the Active Motif kit protocol. Proteins were quantified by Bradford assay (Bio-Rad) according to the manufacturer’s instructions. Proteins were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE). The gels were blotted onto a Nitrocellulose Membrane (Bio-Rad) and blocked with 5% non-fat dry milk and reacted with the appropriate antibodies for BAF subunits: anti-BAF57/SMARCE1 antibody (Abcam ab131328) and anti-SNF5/SMARCB1 antibody (Abcam ab126734), which are rabbit monoclonal against the human subunits and are known to cross-react with zebrafish Smarce1 and Smarcb1 respectively, all membranes were incubated with anti-ERK2 antibody (Santa Cruz Biotechnology, USA; Cat. Nr. sc-153) to confirm equal protein loading. The blots were posteriorly incubated with horseradish peroxide HRP-conjugated anti-mouse IgG or HRP-conjugated anti-rabbit IgG (Santa Cruz Biotechnology). Chemiluminescence was detected with enhanced chemiluminescence (ECL) western blot detection kits (Thermo Scientific Pierce).
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