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Analyses of Bcl-2 (pS70) expression. Panel 1: Western blot analysis of Bcl-2 (pS70) expressed by human Jurkat cells. Lysates (15 µg total cell protein/lane) from untreated (C) and Taxol-treated (T) (Paclitaxel, Sigma, Cat. No. T7191; 100 nM, 24 h) Jurkat cells were blotted using Purified Mouse Anti-Bcl-2 (pS70) antibody (Cat. No. 562529; 0.125, 0.063, and 0.032 µg/ml for Lanes 1, 2, and 3, respectively), HRP Goat Anti-Mouse Ig (Cat. No. 554002) and a chemiluminescent detection system. Panel 2a: Western blot analysis of Bcl-2 (pS70) expressed by human peripheral blood mononuclear cells (PBMC). Phytohemagglutinin-stimulated (PHA, 20 μg/ml for 3 days; Sigma Cat. No. L1668) PBMC were cultured with or without Taxol (100 nM, 24 hr, 37°C). Lysates from 1 million untreated (C) and Taxol-treated (T) PBMC were blotted using Purified Mouse Anti-Bcl-2 (pS70) antibody (2.0 µg/ml) as described above. Panel 2b: Flow cytometric analysis of Bcl-2 (pS70) expressed by human PBMC. PHA-stimulated (20 μg/ml for 3 days) PBMC were cultured with (solid line histogram) or without (dashed line histogram) Taxol (100 nM, 24 hr, 37°C). Cells were fixed in BD Phosflow™ Cytofix Buffer (Cat. No. 554655; 10 min, 37˚C) and permeabilized in BD Phosflow™ Perm Buffer III (Cat. No. 558050; 30 min, on ice) prior to staining with BD Phosflow™ PE Mouse Anti-Bcl-2 (pS70) (Cat. No. 562532) antibody. Fluorescence histograms showing Bcl-2 (pS70) expression were generated for gated events with the forward and side-light scatter characteristics of intact lymphocytes. Panel 3: Immunohistochemical staining for Bcl-2 (pS70). A citrate-pretreated, formalin-fixed, paraffin-embedded tissue section of human tonsil was stained with isotype control (Cat. No. 550878; Left) or Purified Mouse Anti-Bcl-2 (pS70) antibody (Right) (20X original magnification). Note: Bcl-2 (pS70) was identified as a ~28 kDa band in the cell lysates by Western blotting.
BD™ Phosflow Purified Mouse anti-Human Bcl-2 (pS70)
BD™ Phosflow Purified Mouse anti-Human Bcl-2 (pS70)
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The N46-467 monoclonal antibody specifically binds to Bcl-2 (pS70), ie, the Bcl-2 protein phosphorylated at the Ser70 site. Bcl-2 is a ~ 26 kDa intracellular, integral membrane protein found primarily in the nuclear envelope, endoplasmic reticulum and outer mitochondrial membrane. Bcl-2 is encoded by the BCL2 (B-cell CLL/lymphoma 2) gene and is also known as Apoptosis regulator Bcl-2. Members of the Bcl-2 family play a major role in regulating the response of cells to apoptotic signals. Bcl-2 is one of the anti-apoptotic members of the Bcl-2 family. Bcl-2 knockout mice showed pronounced lymphoid apoptosis and other apoptosis related lesions later in life. Bcl-2 is a proto-oncogene because it blocks apoptosis and provides a selective survival advantage in many cell types and thus contributes to tumorigenesis. It has been implicated in several types of cancers, such as breast, prostate, and melanoma . Bcl-2 contains multiple phosphorylation sites including Thr56, Ser70, Thr74 and Ser87. Phosphorylation of Bcl-2 Ser70 has been shown to be a mitotic marker. Phosphorylation at this site regulates Bcl-2's anti-apoptotic activity and has recently been implicated in promoting autophagy. Several studies have shown that Bcl-2 phosphorylation is caused by c-Jun N-terminal kinase (JNK).
研发参考 (6)
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Deng X, Xiao L, Lang W, et al. Novel role for JNK as a stress-activated Bcl2 kinase. J Biol Chem. 2001; 276(26):23681-23688. (Biology). 查看参考
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Geng F, Tang L, Li Y, et al. Allyl isothiocyanate arrests cancer cells in mitosis, and mitotic arrest in turn leads to apoptosis via Bcl-2 protein phosphorylation. J Biol Chem. 2011; 286(37):32259-32267. (Biology). 查看参考
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Ling YH, Tornos C, Perez-Soler R. Phosphorylation of Bcl-2 is a marker of M phase events and not a determinant of apoptosis. J Biol Chem. 1998; 273(30):18984-18991. (Biology). 查看参考
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Maundrell K, Antonsson B, Magnenat E, et al. Bcl-2 undergoes phosphorylation by c-Jun N-terminal kinase/stress-activated protein kinases in the presence of the constitutively active GTP-binding protein Rac1. J Biol Chem. 1997; 272(40):25238-42. (Biology). 查看参考
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Pattingre S, Bauvy C, Carpentier S, Levade T, Levine B, Codogno P.. Role of JNK1-dependent Bcl-2 phosphorylation in ceramide-induced macroautophagy. J Biol Chem. 2009; 284(5):2719-2728. (Biology). 查看参考
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Sarkar S, Korolchuk VI, Renna M, et al. Complex inhibitory effects of nitric oxide on autophagy. Mol Cell. 2011; 43(1):19-32. (Biology). 查看参考
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