Anti myc tag antibody ab9106

Human Type I PDZ GPCR cDNAs were purchased from the Missouri S&T cDNA resource center (cdna.org) or cloned from human brain cDNA library (provided by Professor Ning Zheng, HHMI, University of Washington Department of Pharmacology). Human α-syntrophin cDNA was purchased from OriGene Technologies, (Rockville, MD, USA). Human FLAG-Scribble was provided by Professor Jon Huibregtse (Department of Molecular Sciences, University of Texas at Austin). cDNAs were subcloned into pGlue (provided by Professor Randall T. Moon, HHMI, University of Washington Department of Pharmacology) using In-Fusion HD cloning technology (Clontech).
(R)-(−)-phenylephrine hydrochloride (P6126) was purchased from Sigma (St Louis, MO, USA). Anti-FLAG antibodies were purchased from Sigma (2368). Alexa fluor 633 goat-anti Rabbit IgG (A-21070) and Alexa fluor 568 Goat anti-rat (A-11011) antibodies from Life Technologies. Topro-3 iodide (T3605) is from Life Technologies. Rabbit polyclonal α-syntrophin (H-65, sc-50460), and rabbit polyclonal Scrib (H-300, sc-28737) antibodies from Santa Cruz Biotechnology. Mouse monoclonal Anti-syntrophin (1351, ab11425) antibody and rabbit polyclonal anti-Myc tag antibody (ab9106) from Abcam (Cambridge, MA, USA). Anti-HA mouse mAb (6E2, #2367) from Cell Signaling (Danvers, MA, USA). IRdye 680 goat antimouse IgG and IRdye 800cw goat antirabbit IgG from Li-Cor (Lincoln, NE, USA).

Cells were lysed in lysis buffer containing 50 mM HEPES, 150 mN NaCl (4.38 g), 1 mM EDTA, 1% (w/v) CHAPS and Sigma protease inhibitor cocktail. Subsequently, the cell lysates were boiled in 2 × SDS–PAGE loading buffer for 10 min, and then resolved by 10% SDS–PAGE and transferred to the nitrocellulose membrane. All primary antibodies were diluted 1,000 times for western blotting if not specified. The following antibodies were used in this study: TEAD4 (#ab58310) antibody, anti-Myc tag antibody (#ab9106) and anti-HA tag antibody (#ab9110) were purchased from Abcam; N-cadherin (#610921) antibody was purchase from BD; Vimentin (#5741) and YAP (#12395) antibodies were purchased from Cell Signaling Technology; and alpha-tubulin (#T5168, 1:5000 dilution) and FLAG M2 (#F1804) were purchased from Sigma-Aldrich. RBM4 antibody (#11614-1-AP) was purchased from Proteintech. Bound antibodies were visualized using the ECL kit (GE Healthcare).