Non phospho β cateninser33 37 thr41

Manufactured by Cell Signaling Technology

Non-phospho-β-CateninSer33/37/Thr41 is an antibody that detects β-catenin that is not phosphorylated at Ser33, Ser37, and Thr41. β-catenin is a key component of the Wnt signaling pathway and plays a role in cell-cell adhesion and gene transcription.

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3 protocols using non phospho β cateninser33 37 thr41

Comprehensive Western Blot Analysis

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Western blot analyses were performed as previously described (Jiang et al., 2016 (link); Jiang et al., 2017a ; Jiang et al., 2017b (link)). The HDAC Antibody Sampler Kit (#9928), Phospho-HDAC4^Ser246/HDAC5^Ser259/HDAC7^Ser155 (#3443), HDAC2 (#5113), HDAC6 (#7558), H3K4me2 (#9725), H3K4me3 (#9727), H3K27me2 (#9728), H3K27me3 (#9733), Phospho-AKT^Ser473 (#4060), AKT (#4685), Phospho-mTOR^Ser2448 (#5536), mTOR (#2983), Phospho-AMPKα^Thr172 (#2535), AMPKα (#5831), Phospho-MEK1/2^Ser217/221 (#9154), Phospho-ERK1/2^{Thr202/Tyr204} (#4370), ERK1/2 (#4695), Phospho-JNK1/2 ^{Thr183/Tyr185} (#4668), JNK1/2 (#9258), Phospho-p38^{Thr180/Tyr182} (#4511), p38 (#8690), Phospho-β-Catenin^Ser675 (#4176), Non-phospho-β-Catenin^{Ser33/37/Thr41} (#8814), β-Catenin (#8480), Phospho-IKKα/β^Ser176/180 (#2697), IKKβ (#8943), Phospho-NF-κB p65^Ser536 (#3033), NF-κB p65 (#8242), Phospho-Smad2^Ser465/467 (#3108), Smad2 (#5339), Phospho-Smad3^Ser423/425 (#9520), Smad3 (#9523), Phospho-Smad1/5^Ser463/465 (#9516), and Smad1 (#6944) antibodies were obtained from Cell Signaling Technology. Antibodies against Histone H3 (ab1791), H3K36me2 (ab9049), H3K36me3 (ab9050), H4K20me3 (ab9053), H3K9me1 (ab9045), H3K9me2 (ab1220), H3K4me1 (ab8895), H3K23me1 (ab176132), H4K5ac (ab51997), H4K8ac (ab15823), H3K18ac (ab40888), H3K23ac (ab61234) and H4K12ac (ab46983) were purchased from Abcam.

Guo X., Fang Z.M., Wei X., Huo B., Yi X., Cheng C., Chen J., Zhu X.H., Bokha A.O, & Jiang D.S. (2019). HDAC6 is associated with the formation of aortic dissection in human. Molecular Medicine, 25, 10.

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EP Receptor and β-Catenin Expression

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Protein samples were loaded equally based on concentration, fractionated using polyacrylamide gel electrophoresis (PAGE), and transferred onto a nitrocellulose membrane. EP receptors’ protein expression was tested using the primary rabbit polyclonal antibodies anti-EP1 (Santa Cruz Biotechnology), anti-EP2 (Santa Cruz Biotechnology), anti-EP3 (Santa Cruz Biotechnology), and anti-EP4 (Santa Cruz Biotechnology). The anti-EP3 antiody recognizes all three isoforms (α, β, γ) of the EP3 protein. The levels of β-catenin were tested using the primary antibodies against phospho-β-catenin (Ser552) and non-phospho β-catenin (Ser33/37/Thr41) (Cell Signaling), respectively. Rabbit polyclonal anti-phospho-β-catenin (Ser552) antibody recognizes the active form phosphorylated by protein kinase A (PKA), and rabbit monoclonal anti-nonphospho-β-catenin (Ser33/37/Thr41) antibody recognizes the stabilized form of β-catenin that is functionally active. For comparative analysis, anti-GAPDH (Abcam) was used on the same blots. Horseradish peroxidase-conjugated secondary antibodies used were α-rabbit (Abcam), α-goat (Santa Cruz Biotechnology), or α-mouse (Abcam). Clarity Western ECL Substrate reagent (Bio-rad) was used for detection by Perkin Elmer Geliance 600 Imaging System.

Rai-Bhogal R., Wong C., Kissoondoyal A., Davidson J., Li H, & Crawford D.A. (2018). Maternal exposure to prostaglandin E2 modifies expression of Wnt genes in mouse brain – An autism connection. Biochemistry and Biophysics Reports, 14, 43-53.

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Western Blot Analysis of Tumor Samples

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Western blot analysis was performed as previously described43 . Whole protein lysates of tumor samples were collected by the abrading of tumors in liquid nitrogen. All the samples were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and probed overnight at 4 °C with the following primary antibodies: FLT3 (1:500, Santa Cruz Biotechnology, sc-480), Phospho-FLT3(Tyr589/591) (1:1000, Cell Signaling, #3464), STAT5 (1:1000, Cell Signaling, #9363), Phospho-STAT5(Tyr694) (1:1000, Cell Signaling, #9314), p44/42 MAPK (1:1000, Cell Signaling, #4695), Phospho-p44/42 MAPK(Thr 202/Tyr204) (1:1000, Cell Signaling, #4370), Non-phospho β-catenin (Ser33/37/Thr41) (1:1000, Cell Signaling, #8814), β-catenin (1:500, Santa Cruz Biotechnology, sc-59737), NKD1 (1:10000, abcam, ab133650) and β-actin (1:1000, ZSGB-BIO, TA-09). Quantitative analysis of the protein level was performed with NIH image analysis software, ImageJ (normalized relative to the vehicle control).

Ma S., Yang L.L., Niu T., Cheng C., Zhong L., Zheng M.W., Xiong Y., Li L.L., Xiang R., Chen L.J., Zhou Q., Wei Y.Q, & Yang S.Y. (2015). SKLB-677, an FLT3 and Wnt/β-catenin signaling inhibitor, displays potent activity in models of FLT3-driven AML. Scientific Reports, 5, 15646.

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