Pbcl 2 ser70

Manufactured by Cell Signaling Technology

Sourced in United States

PBcl-2(Ser70) is a lab equipment product designed for use in scientific research. It is a phospho-specific antibody that recognizes the Ser70 phosphorylation site on the Bcl-2 protein. The core function of this product is to enable the detection and analysis of Bcl-2 phosphorylation, which is a key regulatory mechanism in cellular processes.

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Lab products found in correlation

4 protocols using pbcl 2 ser70

Synthesis and Evaluation of SKI-178 Compound

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SKI-178
(N′-[(1E)-1-(3,4-dimethoxyphenyl)ethylidene]-3-(4-methoxxyphenyl)-1H-pyrazole-5-carbohydrazide)
was synthesized as described previously.^{24 (link)} Reagents were purchased as follows:vincristine (Thermo
Fisher Scientific, Waltham, MA, USA), Colchicine (Enzo Life Sciences,
Farmingdale, NY, USA), Paclitaxel (Sigma-Aldrich, St. Louis, MO, USA), MG132
(Sigma-Aldrich, St. Louis, MO, USA) and PF-543 (Selleck Chemicals, Houston, TX,
USA), antibodies against SphK1, pBcl-2(Ser70), Cleaved caspase-7, and Tubulin
(Cell Signaling Technology, Danvers, MA, USA), SphK1 (pSer225) (ECM Biosciences,
Versailles, KY), and GAPDH (Santa Cruz Biotechnology, Santa Cruz, CA, USA).
β-hydroxy-propyl-cyclodextrin was from Acros (NJ, USA). 1,2-Propanediol,
Tween-20, and dextrose were from Sigma Aldrich (MO, USA).

Hengst J.A., Dick T.E., Sharma A., Doi K., Hegde S., Tan S.F., Geffert L.M., Fox T.E., Sharma A.K., Desai D., Amin S., Kester M., Loughran T.P., Paulson R.F., Claxton D.F., Wang H.G, & Yun J.K. (2017). SKI-178: A Multitargeted Inhibitor of Sphingosine Kinase and Microtubule Dynamics Demonstrating Therapeutic Efficacy in Acute Myeloid Leukemia Models. Cancer translational medicine, 3(4), 109-121.

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Western Blotting of Cell Signaling Proteins

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Cell protein lysates were prepared, and Western blotting was performed according to our published protocols [34 (link), 105 (link), 107 (link)]. The following primary antibodies were used in this experiment: E2F1 (3742, Cell Signaling), E2F2 (sc-633, Santa Cruz Biotechnology), E2F3 (sc-878, Santa Cruz Biotechnology), cyclin B1 (sc-245, Santa Cruz Biotechnology), Mad2 (ab70383, Abcam), Bcl2 (2870, Cell Signaling), pBcl2 (Ser70) (2827, Cell Signaling), pSer/Thr/Phe (9631, Cell Signaling), cleaved caspase 3 (9661, Cell Signaling), cleaved PARP (5625, Cell Signaling), Hec1 (GTX70268, GeneTex, Irvine, CA), pBad (Ser136) (4366, Cell Signaling), Bad (9268, Cell Signaling), Mps1/TTK (3255, Cell Signaling), PP2Ac (2259, Cell Signaling), and Sgo1 (ab58023, Abcam). β-actin antibody (4970, Cell Signaling) was used as a loading control. For secondary antibodies, either goat anti-rabbit HRP (sc-2004) or goat anti-mouse HRP (sc-2005, Santa Cruz Biotechnology) were used. Signals were detected by using a Lumigen TMA-6 reagent (Lumigen Inc, Southfield, MI). Image J software (NIH, Bethesda, MD) was used to quantify protein levels.

Lee M., Rivera-Rivera Y., Moreno C.S, & Saavedra H.I. (2017). The E2F activators control multiple mitotic regulators and maintain genomic integrity through Sgo1 and BubR1. Oncotarget, 8(44), 77649-77672.

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Western Blot Analysis of Cellular Signaling

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Western blot analysis was performed as previously described.12 PVDF membranes (Millipore, Billerica, MA) were primed with primary antibodies against VEGFR1 (vascular endothelial growth factor receptor 1), VEGFR2 (vascular endothelial growth factor receptor 1), ERK1/2 (extracellular signaling kinase 1 and 2), eNOS (endothelial nitric oxide synthase), p‐FOXO1 (phosphorylated forkhead box O1), AMPK‐α (AMP‐activated protein kinase‐α), FOXO1 (forkhead box O1), BAD (B‐cell lymphoma 2 [Bcl‐2]‐associated death promotor), SOD2 (superoxide dismutase 2), p‐IRS‐1 Ser332 (phosphorylated insulin receptor substrate 1 serine 332), p‐Bcl‐2 ser70 (phosphorylated Bcl‐2 serine 70) and p‐mcl‐1 ser159 (phosphorylated myeloid leukemia cell differentiation protein 1 serine 159; all from Cell Signaling Technology, Danvers, MA). ImageJ software (NIH, Bethesda, MD) was used to quantify band densitometry as arbitrary light units. Loading error was controlled for by probing membranes with an antibody against GAPDH.

Potz B.A., Sabe A.A., Elmadhun N.Y., Clements R.T., Robich M.P., Sodha N.R, & Sellke F.W. (2016). Glycogen Synthase Kinase 3β Inhibition Improves Myocardial Angiogenesis and Perfusion in a Swine Model of Metabolic Syndrome. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 5(7), e003694.

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Tetrandrine-Induced Apoptosis in Cancer

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Tetrandrine was purchased from Enzo Life Sciences (Farmingdale, NY). The cell fractionation kit was purchased from MitoScience Inc. (Eugene, OR), protein A/G-agarose from Santa Cruz Biotechnology (Santa Cruz, CA), and MG132 and z-DEVD-FMK from Cayman Chemical (Ann Arbor, MI). Antibodies against Bax, Bcl₂, Apaf-1, cytochrome c, c-Jun, Fas, FADD, FasL, c-FLIP_L/S and GAPDH were obtained from Santa Cruz Biotechnology (Santa Cruz, CA), whereas ubiquitin, PARP, Bid, p-JNK Thr183/Tyr185, JNK1/2, p-Bcl₂ Ser70, p-ASK1 Thr845, ASK1, p-SEK1 Thr261, SEK1, p-c-Jun Ser63, Smac and caspase-3, -8 and -9 were from Cell Signaling Technology (Danvers, MA). The cell culture medium RPMI-1640 and fetal bovine serum were from GIBCO (Invitrogen, Carlsbad, CA). All other reagents and chemicals were purchased from Sigma-Aldrich (St. Louis, MO).

Chaudhary P, & Vishwanatha J.K. (2014). c-Jun NH2-terminal kinase-induced proteasomal degradation of c-FLIPL/S and Bcl2 sensitize prostate cancer cells to Fas- and mitochondria-mediated apoptosis by tetrandrine. Biochemical pharmacology, 91(4), 457-473.

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