Total proteins were extracted in RIPA lysis and extraction buffer supplemented with 1% protease inhibitor cocktail (Roche, Indianapolis, IN, USA). The protein levels were measured using the Bradford protein assay and 15 μg of proteins were loaded onto an SDS-PAGE gel and separated before transfer onto a nitrocellulose membrane. The blots were probed using mouse monoclonal anti-KCa1.1 (clone L6/60; UC Davis/NIH Neuromab Facility) or rabbit polyclonal anti-NFκB p65 (Santa Cruz Biotechnology, Dallas, TX, USA) antibodies and rabbit monoclonal antibody against β-actin (Sigma). The blots were further incubated with secondary antibodies conjugated to fluorophores (LI-COR, Lincoln, Nebraska) and visualized using an Odyssey Imaging System (LI-COR).
Rabbit monoclonal antibody against β actin
Manufactured by Merck Group
Sourced in United States
The Rabbit monoclonal antibody against β-actin is a laboratory reagent used for the detection and quantification of the β-actin protein in various biological samples. β-actin is a highly conserved cytoskeletal protein found in all eukaryotic cells and is commonly used as a loading control or reference in various experimental techniques, such as Western blotting, immunohistochemistry, and flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Secondary antibodies conjugated to fluorophores, by LI COR (1 mentions)
Anti nf κb p65, by Santa Cruz Biotechnology (1 mentions)
Odyssey imaging system, by LI COR (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Santa Cruz Biotechnology (1 mentions)
Polyvinylidene difluoride membrane, by Merck Group (1 mentions)
2 protocols using rabbit monoclonal antibody against β actin
1
Western Blot Analysis of Protein Targets
2
Western Blot Analysis of STARD8 Protein
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Whole cell lysates were prepared from GC cell lines. Samples were loaded onto 10% sodium dodecyl sulfate (SDS) poly-acrylamide gels, transferred to polyvinylidene difluoride membranes (EMD Millipore, Billerica, MA, USA) and incubated with primary mouse polyclonal antibody against human STARD8 (Abcam, Cambridge, MA, USA) and rabbit monoclonal antibody against β-actin (Sigma-Aldrich Co., St Louis, MO, USA) overnight at 4°C. Membranes were incubated with a horseradish-peroxidase-conjugated secondary antibody (Santa Cruz Biotechnology Inc., Dallas, TX, USA) for 2 h at room temperature, and immunoreactive protein bands were visualized with a chemiluminescent detection kit (enhanced chemiluminescence [ECL]; Thermol Biotech Inc., Rockford, IL, USA). Signal intensities were quantified and analyzed as described in the following sections. Each experiment was repeated three times.
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