Rabbit anti phospho p53 ser15

Manufactured by Cell Signaling Technology

Sourced in United States

Rabbit anti-phospho-p53 (Ser15) is a primary antibody that specifically recognizes the p53 protein when phosphorylated at serine 15. This antibody is commonly used in research applications to detect and analyze the activation of the p53 signaling pathway.

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Anti-p53 (256 citations) Phospho-p53 (Ser15) (91 citations) Phospho-p53 (57 citations) Anti-phospho-p53 (Ser15) (40 citations) Anti-phospho-p53 (35 citations) Rabbit anti-p53 (23 citations) Ser15-p53 (8 citations) Rabbit anti-phospho-p53 (2 citations)

8 protocols using rabbit anti phospho p53 ser15

Immunofluorescence Analysis of CSFV-Infected Cells

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CSFV-infected cells were washed in phosphate-buffered saline (PBS) and fixed with methanol/acetone (1:1) for 20 min at 25°C ± 2°C and permeabilized with 1% Triton X-100 in PBS for 10 min. After three washes in PBS, samples were incubated with mouse anti-E2 antibody (MssBio, Guangzhou, China) and rabbit anti-phospho-p53(Ser15) (Cell Signaling Technology) for 1 h at 25°C ± 2°C, followed by staining with donkey anti-rabbit IgG conjugated to Alexa Fluor® 594 and donkey anti-mouse IgG conjugated to Alexa Fluor® 488 (YEASEN, Shanghai, China) at a 1:200 dilution for 1 h at 25°C ± 2°C. After incubation with 4′, 6-diamidino-2-phenylindole (DAPI), the samples were observed using a laser-scanning confocal microscope (LSM 510 META; Carl Zeiss, Jena, Germany).

Ning P., Hu C., Li X., Zhou Y., Hu A., Zhang Y., Gao L., Gong C., Guo K., Zhang X, & Zhang Y. (2017). Classical swine fever virus Shimen infection increases p53 signaling to promote cell cycle arrest in porcine alveolar macrophages. Oncotarget, 8(34), 55938-55949.

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Western Blot Analysis of Cell Signaling

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Whole cell lysates for Western Blot were prepared with the SDS-sample buffer. Proteins were separated by SDS-PAGE and transferred onto the PVDF membrane (Millipore/Fisher Scientific, Pittsburg, PA), Darmstadt, Germany). Antibodies for immunoblotting from Cell Signaling Technology (Dancers, MA) would include rabbit anti-Bmal1 (#14020), rabbit anti-CLOCK (#5157), rabbit anti-phospho-p53 (Ser15) [#9284], rabbit anti-p21 Waf1/Cip1 (#2947), rabbit anti-Histone H2AX (#2595), rabbit anti-cleaved PARP (#9541), and rabbit anti-phospho-CHK1 (Ser 345) (#2348). Other antibodies include mouse anti-p53 (sc-126; Santa Cruz Biotechnology Inc., Santa Cruz, CA), rabbit anti-Keratin 10 (Poly19054; Biolegend, San Diego, CA), mouse anti-phospho-Histone H2AX (Ser139) (05–636; Millipore/Fisher Scientific), and mouse anti-α-tubulin (T9026; Sigma-Aldrich, St. Louis, MO). HRP-conjugated secondary antibodies were from Santa Cruz Biotechnology Inc. Chemiluminescence images were acquired using an Amersham Imager 600 from GE Healthcare Life Sciences (Pittsburgh, PA) or iBright FL1000 (Invitrogen-Life Technology/Fisher Scientific, Pittsburgh, PA). The level of target proteins was quantified by densitometry scanning with the ImageJ software and normalized to the amount of α-tubulin.

Sun Y., Wang P., Li H, & Dai J. (2018). BMAL1 and CLOCK proteins in regulating UVB-induced apoptosis and DNA damage responses in human keratinocytes. Journal of cellular physiology, 233(12), 9563-9574.

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Western Blotting of EMT Markers

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Western blotting was performed as described previously using whole cell lysates [14 (link)]. Cells were harvested to prepare cell lysates two weeks after initiating of mutant KRAS^V12 induction. Primary antibodies used were mouse anti-E-cadherin (Cat#610181, BD Biosciences), mouse anti-Vimentin Cat#550513, BD Biosciences), mouse anti-p53 (Cat#sc-126, Santa Cruz, USA), mouse anti-p21 Waf/Cip/CDKN1A (Cat#sc-6246, Santa Cruz), mouse anti-KRAS (Cat#sc-30, Santa Cruz), rabbit anti-AKT (pan) (Cat#4691, Cell Signaling Technology), rabbit anti-phospho-AKT (ser 473) (Cat#4060, Cell Signaling Technology), rabbit anti-p44/42MAPK(Erk1/2) (Cat#9102, Cell Signaling Technology), rabbit anti-phospho-p44/42MAPK(Erk1/2) (Cat#4370, Cell Signaling Technology), mouse anti-Actin (Cat#A2228, Sigma-Aldrich), mouse anti-Vinculin (Cat#SC-73614, Santa Cruz), rabbit anti-α-tubulin (Cat# PM054-7, MBL Life Science, USA), and rabbit anti-phospho-p53 (Ser 15) (Cat#9284, Cell Signaling Technology) antibodies. Actin, Vinculin, or α-tubulin protein levels were used as a control for the adequacy of equal protein loading. Anti-rabbit or anti-mouse (GE Healthcare, Buckinghamshire, England) antibodies were used at 1:2000 dilution as secondary antibodies.

Bisgaard H, & Møller H. (1999). Changes in risk of hospital readmission among asthmatic children in Denmark, 1978-93. BMJ : British Medical Journal, 319(7204), 229-230.

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Western Blot Analysis of Cellular Proteins

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Cells were lysed and extracts prepared as described (24). Primary antibodies included: rabbit anti-PAI-1 (1:3000) [25 (link)], rabbit anti-p53 (1:2000; Cell Signaling), rabbit anti-ERK2 (1:5000; Santa Cruz); rabbit anti-phospho-SMAD2/3 (1:500; Santa Cruz), rabbit anti-β-actin (1:5000; Santa Cruz), mouse anti-Fibronectin (1:2000; BD Biosciences), rabbit anti-CTGF/CCN2 (1:2000, Abcam), mouse anti-α-smooth muscle actin (1:2000; BD Biosciences), rabbit anti-COX-2 (1:2000, Santa Cruz), rabbit anti-p21 (1:1000, Santa Cruz), rabbit anti-phospho-p53 sampler kit (1:1000; Cell Signaling), rabbit anti-phospho-p53^Ser15 (1:1000; Cell Signaling), rabbit anti-phospho-SMAD3 (1:2000, Thermoscientfic), rabbit anti-lamin A C (1:1000, Santa Cruz), rabbit anti-SMAD3 (1:2000, Abcam), rabbit anti-SMAD2 (1:1000, Santa Cruz), mouse anti-GFP (1:2000, Santa Cruz), rabbit anti-acetyl-lysine (1:1000, Cell Signaling), rabbit anti-p300 (1:2000, Santa Cruz).

Overstreet J.M., Samarakoon R., Meldrum K.K, & Higgins P.J. (2014). Redox control of p53 in the transcriptional regulation of TGF-β1 target genes through SMAD cooperativity. Cellular signalling, 26(7), 1427-1436.

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Antibody Characterization for Research

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The following antibodies were used: mouse Anti-p53 (Ab6) (Calbiochem OP43); rabbit anti-phospho-p53 (Ser15) (Cell Signaling 9284); mouse anti-phospho-Histone H2A.X (Ser139) (Millipore 05-636); rabbit anti-Histone H2A.X (Millipore 07-627); rabbit anti-acetyl-Histone H3 (Lys9) (Millipore 07-352); rabbit anti-Histone H3 (K4me1) (abcam ab8895); rabbit anti-Histone H3 (K27Ac) (abcam ab4729); rabbit anti-Histone H3, CT, pan (Millipore 07-690); rabbit anti-GAPDH (14C10) (Cell Signaling 2118).

Tutton S., Deng Z., Gulve N., Vladimirova O., Beishline K., Wiedmer A., Murphy M, & Lieberman P.M. (2019). Elevated telomere dysfunction in cells containing the African-centric Pro47Ser cancer-risk variant of TP53. Oncotarget, 10(38), 3581-3591.

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Western Blot Antibody Panel

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Antibodies used in western blotting: rabbit-anti-phospho-p53 (Ser15) (Cell Signaling Technology, Danvers, USA); mouse-anti-p53 (Santa Cruz Biotechnology, Dallas, USA); rabbit-anti-phospho-p70S6K (Thr389) (Cell Signaling Technology); rabbit-anti-p70S6K (Cell Signaling Technology); mouse-anti-phospho-4EBP1 (Ser65) (Santa Cruz Biotechnology); rabbit-anti-4EBP1 (Cell Signaling Technology); mouse-anti-p62/ SQSTM1 (Santa Cruz Biotechnology); rabbit-anti-LC3B (Thermo Fisher Scientific, Waltham, USA); mouse-anti-β-actin (Santa Cruz Biotechnology); horseradish peroxidase (HRP) conjugated secondary antibodies (Santa Cruz Biotechnology).

Zhang X., Rodriguez-Niño A., Pastene D.O., Pallavi P., van den Born J., Bakker S.J., Krämer B.K, & Yard B.A. (2021). Methylglyoxal induces p53 activation and inhibits mTORC1 in human umbilical vein endothelial cells. Scientific Reports, 11, 8004.

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Analyzing p53 Expression in Fetal Liver

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Western blotting was conducted with fetal liver cells enriched through discontinuous albumin density gradient centrifugation using rabbit anti-p53 (Santa Cruz Biotechnology, Inc.), or rabbit anti–phospho-p53 (Ser15; Cell Signaling Technology) antibodies.

Nishimura S., Nagasaki M., Kunishima S., Sawaguchi A., Sakata A., Sakaguchi H., Ohmori T., Manabe I., Italiano JE J.r., Ryu T., Takayama N., Komuro I., Kadowaki T., Eto K, & Nagai R. (2015). IL-1α induces thrombopoiesis through megakaryocyte rupture in response to acute platelet needs. The Journal of Cell Biology, 209(3), 453-466.

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Western Blot for Phospho-p53 and Lamin A/C

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Western blot analysis was performed as reported previously (21) (link). Phospho-p53 (Ser15), p53, lamin A/C, and α-tubulin were detected using antibodies against rabbit anti-phospho-p53 (Ser15) (1:250 dilution, Cell Signaling Technology, Inc), mouse anti-p53 (1:2000 dilution, Merck), Lamin A+C (1:3000 dilution, Gene Tex), and alpha Tubulin 4a (1:5000 dilution, GeneTex), respectively.

Yamamoto A., Kawami M., Konaka T., Takenaka S., Yumoto R, & Takano M. (2019). Anticancer Drug-Induced Epithelial-Mesenchymal Transition via p53/miR-34a axis in A549/ABCA3 Cells. Journal of pharmacy & pharmaceutical sciences : a publication of the Canadian Society for Pharmaceutical Sciences, Societe canadienne des sciences pharmaceutiques, 22(1).

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