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2 protocols using grk5 c 20

1

Western Blot Analysis of Cellular Signaling Proteins

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Protein was extracted, quantified, and probed with antibody, as previously described.69 (link) Whole cell lysates were extracted using RIPA buffer (Cell Signaling). Nuclear and cytoplasmic lysates were extracted using the NE-PER Nuclear and Cytoplasmic Extraction Kit, according to the manufacturer (Thermo Scientific). Antibodies included AKT (Cell Signaling, Danvers, MA), phospho-AKT (S473) (Cell Signaling), CXCR4 (Abcam, Cambridge, MA), phospho-CXCR4 (S339) (Abcam), eEF2 (Cell Signaling), GRK4-6 (EMD Millipore, Billerica, MA), GRK5 (C-20) (Santa Cruz Biotechnology, Inc., Santa Cruz, CA), KCNA1 (Abcam), Lamin B1 (Cell Signaling), NPR3 (Abcam), WIP1 (Bethyl, Montgomery, TX), and β-actin (Sigma-Aldrich, St. Louis, MO). Secondary antibodies Alexa Fluor 680 goat anti-mouse IgG (Life Technologies) or IRDye 800 goat anti-rabbit IgG (Rockland, Gilbertsville, PA) were used at a dilution of 1:5,000. Immunoblots were imaged and quantified, as previously described.27 (link)
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2

Western Blot Analysis of Cellular Signaling Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
Protein was extracted, quantified, and probed with antibody, as previously described.69 (link) Whole cell lysates were extracted using RIPA buffer (Cell Signaling). Nuclear and cytoplasmic lysates were extracted using the NE-PER Nuclear and Cytoplasmic Extraction Kit, according to the manufacturer (Thermo Scientific). Antibodies included AKT (Cell Signaling, Danvers, MA), phospho-AKT (S473) (Cell Signaling), CXCR4 (Abcam, Cambridge, MA), phospho-CXCR4 (S339) (Abcam), eEF2 (Cell Signaling), GRK4-6 (EMD Millipore, Billerica, MA), GRK5 (C-20) (Santa Cruz Biotechnology, Inc., Santa Cruz, CA), KCNA1 (Abcam), Lamin B1 (Cell Signaling), NPR3 (Abcam), WIP1 (Bethyl, Montgomery, TX), and β-actin (Sigma-Aldrich, St. Louis, MO). Secondary antibodies Alexa Fluor 680 goat anti-mouse IgG (Life Technologies) or IRDye 800 goat anti-rabbit IgG (Rockland, Gilbertsville, PA) were used at a dilution of 1:5,000. Immunoblots were imaged and quantified, as previously described.27 (link)
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