p300 (D2X6N) Rabbit mAb 54062S from Cell Signaling Technology

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Nature communications
April 27 2022

Fig 1: wt ATF1 is present at a majority of EWSR1-ATF1 binding sites.a Heatmaps depicting EWSR1 N, ATF1 C and ATF1 N ChIP-seq signal intensities at 2011 distal and 374 proximal EWSR1-ATF1 binding sites in SU-CCS-1 and primary CCS1 tumor cells, illustrating genomic co-occupancy by the wt ATF1 TF at the fusion protein binding sites. For each heatmap 20 kb regions centered on the EWSR1-ATF1 peaks are shown. Signals are ranked by ATF1 C intensity. b Composite plots showing changes in ChIP-seq signal for ATF1 N peaks associated (left) or not (right) with EWSR1-ATF1 binding in SU-CCS-1 cells transfected with a siRNA targeting the fusion gene (siEA) or control (siCTL) siRNAs. The loss of the fusion protein is associated with a decrease in ATF1 N signal only at EWSR1-ATF1 bound sites, suggesting wt ATF1 recruitment by the fusion protein. c Co-immunoprecipitation (Co-IP) assay showing the direct interaction between the EWSR1-ATF1 and wt ATF1 proteins in 293 T (left) and DTC1 cells (right). 293 T cells were transfected with either a V5-tagged EWSR1-ATF1 (EA-CV5) or empty vector (CTL) constructs. The IP was performed using an anti-V5 tag antibody, and the western blot revealed using anti-V5 (top) or anti-ATF1 C (bottom) antibodies. DTC1 IPs were performed using anti-ATF1 C or -ATF1 N antibodies, compared to control (CTL) IgG, and the western blot revealed using an anti-ATF1 C antibody. The experiment was independently repeated twice with similar results. d Correlation plot of EWSR1 N, ATF1 N, and H3K27ac ChIP-seq scores at the 2011 distal regions, showing stronger correlation between H3K27ac deposition and wt ATF1 presence, than with EWSR1-ATF1 presence (represented by ATF1 N and EWSR1 N signals, respectively). e Scatter plots of ATF1 N and p300 (left), or EWSR1 N and p300 (right) ChIP-seq scores, illustrating that wt ATF1 presence correlates better with p300 occupancy than EWSR1-ATF1. f Western blot analysis of EWSR1-ATF1 and wt ATF1 protein levels in DTC1 and SU-CCS-1 cell lines infected with a shRNA targeting the wt ATF1 transcript (shATF1) or an unrelated sequence (shCTL) at 72 h post-lentiviral infection. Tubulin protein levels were used as loading control. The experiment was independently repeated twice with similar results. g qPCR analysis showing reduction of wt ATF1 transcripts, as well as a panel of EWSR1-ATF1 / wt ATF1 co-regulated target transcripts, in shATF1- vs shCTL-infected SU-CCS-1 (S) and DTC1 (D) cells at 72 h post-lentiviral infection. Three replicates of each condition were used to calculate mean Ct values, mean relative expression values were then calculated according to the 2-??Ct method relative to the shCTL sample values, and normalized to the endogenous control gene GAPDH. Error bars show standard deviation of mean values (n = 3 sample replicates). The experiment was independently repeated twice with similar results. Source data are provided in the Source Data file.
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Cell death & disease
October 29 2022

Fig 2: BRD4 promotes the transcription levels of HADH, ACSL1 and ACAA2 genes.A The enrichment of BRD4, H3K27ac, P300, and Pol II in the promoter region of HADH, ACSL1, and ACAA2 via the ChIP/ qRT-PCR assay. Transcription activity of BRD4 on HADH, ACSL1, and ACAA2 promoters regions via dual-luciferase assay (B) and yeast one-hybrid system C. (n = 3, Mean±SD, one-way ANOVA, *P < 0.05; **P < 0.01; ***P < 0.005; N.S.: not significant). The mRNA (D) and protein (E) expression levels of HADH, ACSL1, and ACAA2 detected by qRT-PCR and Western blotting (n = 3, Mean ± SD, one-way ANOVA, *P < 0.05; **P < 0.01; ***P < 0.005; N.S.: no significant). F Co-localization analysis between HADH/ ACSL1/ ACAA2 and the mitochondria.
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Oncogenesis
October 27 2021

Fig 3: Inhibition of HIF-1α or Smad3 could reduce cell viability and self-renewal in the proneural and mesenchymal subtypes of GSCs.A Heatmap showing molecular subtype marker expression in proneural GSCs (PN12, PN16, and PN19), mesenchymal GSCs (ME23, ME27, and ME29), and proneural (or mesenchymal) GSCs infected with the indicated plasmids or shRNA or treated with the indicated inhibitors. HIF-1α inhibitor, GN44028, 12.5 μM. Palmitoylation inhibitor, 2-BP, 100 μM. Smad3 inhibitor, (E)-SIS3, 20 μM. EP300 inhibitor, C646, 15 μM. Z-scores were calculated from the △Ct values obtained in the quantitative polymerase chain reaction analysis. B Viability of proneural PN12 GSCs transfected with the indicated plasmids or shRNA or treated with GN44028, C646, 2-BP, or (E)-SIS3 for 48 h. C A serial dilution assay was used to determine the self-renewal capacity of PN12 GSCs transfected with the indicated plasmids or shRNA or treated with GN44028, C646, 2-BP, or (E)-SIS3. D Viability of mesenchymal ME23 GSCs transfected with the indicated plasmids or shRNA or treated with GN44028, C646, 2-BP, or (E)-SIS3 for 48 h. E A serial dilution assay was used to determine the self-renewal capacity of ME23 GSCs transfected with the indicated plasmids or shRNA or treated with GN44028, C646, 2-BP, or (E)-SIS3. GSCs glioblastoma stem cells, HIF-1α hypoxia-inducible factor 1-alpha.
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Journal of Biological Chemistry
December 01 2021

Fig 4: Identification of the SRC family coactivators as Sox2-interacting proteins.A, GST-tagged c-Myc, Klf4, Oct4, and Sox2 were used to pulldown 35S-methionine-labeled coregulators synthesized via in vitro–coupled transcription and translation reactions. The bound coregulators were subsequently visualized by autoradiography. B, co-IP assay confirming the interaction between Sox2 and SRC family coactivators. The plasmids encoding FLAG-Sox2 and Myc-SRCs were cotransfected into HEK293T cells as indicated, and the interaction between Sox2 and each SRC proteins was determined by immunoprecipitation of Myc-SRCs followed by WB analysis using the anti-FLAG antibody. C, co-IP assays using mouse E14 ES cell extracts were performed to detect the interaction of endogenous Sox2 with endogenous SRC-1, SRC-2, SRC-3, and p300. D and E, ChIP assays were performed by analyzing the association of Sox2 (D) and p300, SRC-1, SRC-2, and SRC-3 (E) with five Sox2 target genes. The ChIP data were relative to the percentage of input. ChIP, chromatin immunoprecipitation; co-IP, co-immunoprecipitation; ES, embryonic stem; Sox2, SRY-box 2; SRCs, steroid receptor coactivators.
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Journal of Biological Chemistry
December 01 2021

Fig 5: SRC family coactivators enhance Sox2 transcriptional activity.A–C, luciferase reporter assay showing that the SRC family coactivators SRC-1 (A), SRC-2 (B), and SRC-3 (C) enhanced transcriptional activation by Sox2 in a dose-dependent manner. Also shown are WB results of ectopically expressed FLAG-Sox2 with Myc-SRC-1, Myc-SRC-2, or Myc-SRC-3, respectively. GAPDH serves as a loading control. FGF4-Luc reporter, 50 ng; FLAG-Sox2, 200 ng; Myc-SRC-1/2/3, 200 ng, 500 ng, and 1000 ng for an increasing dose of SRC expression plasmids. D–F, luciferase reporter assay showing that SRC-1 (D), SRC-2 (E), and SRC-3 (F) promoted Sox2 transcriptional activity synergistically with p300. Also shown are WB results of ectopically expressed FLAG-Sox2, Myc-p300, and Myc-SRC-1, Myc-SRC-2, or Myc-SRC-3, respectively. FGF4-Luc, 50 ng; FLAG-Sox2, 200 ng; Myc-SRC-1/2/3, 200 ng and 500 ng; Myc-p300, 1000 ng. Data are represented as relative luciferase activity (×104) (mean ± SD of three experimental repeats). All statistical analyses were performed using Excel 2016. *p< 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. Sox2, SRY-box 2; SRCs, steroid receptor coactivators.

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Cell Signaling Technology for
p300 (D2X6N) Rabbit mAb

The p300 (D2X6N) Rabbit mAb Antibody from Cell Signaling Technology is a Rabbit Monoclonal antibody to E1A binding protein p300, and EP300. This antibody recognizes Homo Sapiens, and Human antigen. The p300 (D2X6N) Rabbit mAb Antibody has been validated for the following applications: Precipitation, ChIP, Immunoprecipitation, Western Blot, and Other.

Description

p300 (D2X6N) Rabbit mAb recognizes endogenous levels of total p300 protein. This antibody does not cross-react with CBP protein

Product Specs
Itemp300 (D2X6N) Rabbit mAb
CompanyCell Signaling Technology
Price Pricing Info Supplier Page View Company Product Page
Catalog Number54062S
Quantity100 µl
ApplicationsW, IP, ChIP, C&R
ReactivityH
Targetp300
IsotypeIgG
HostRabbit
NCBI Full Gene NameE1A binding protein p300
NCBI Gene AliasesKAT3B, MKHK2, RSTS2, p300
ClonalityMonoclonal
Research AreaChromatin Regulation / Nuclear Function
Molecular Weight300

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Website: Technical Support

(1) Targeting c-Jun inhibits fatty acid oxidation to overcome tamoxifen resistance in estrogen receptor-positive breast cancerCell Death & DiseaseOctober 6, 2023Cen Jiang, Youzhi Zhu, Huaying Chen, Junyu Lin, Ruiwang Xie, Weiwei Li, Jiajie Xue, Ling Chen, Xiangjin Chen, Sunwang XuCPT1C (12969-1-AP, dilution 1:500) was purchased from Proteintech, CBP (#7389, dilution 1:1000), P300 (#54062, dilution 1:1000), H3 (#4499, dilution 1:2000) were purchased from Cell Signaling Tech, β-actin (A1978,

(2) A chemical catalyst enabling histone acylation with endogenous acyl-CoANature CommunicationsSeptember 22, 2023Misuzu Habazaki, Shinsuke Mizumoto, Hidetoshi Kajino, Tomoya Kujirai, Hitoshi Kurumizaka, Shigehiro A. Kawashima, Kenzo Yamatsugu, Motomu KanaiPTM-901, 1:1000 dilution for WB), a rabbit monoclonal antibody against p300 (Cell Signaling Technology, 54062, 1:1000 dilution for WB), and mouse monoclonal antibodies against H2BK120ac (1 μg/mL for WB) and H3K27ac

(3) KLF12 transcriptionally regulates PD‐L1 expression in non‐small cell lung cancerMolecular OncologySeptember 2, 2023Xiaohui Pan, Wenxin Zhang, Longsheng Wang, Hongjie Guo, Mingming Zheng, Honghai Wu, Qinjie Weng, Qiaojun He, Ling Ding, Bo Yang(Cambridge, UK). Anti‐PD‐L1 (#13684S), Anti‐Phospho‐STAT1 (#7649), Anti‐Phospho‐STAT3 (#7649), Anti‐P300 (#54062), and Anti‐CBP (#7389) were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti‐β‐actin

(4) Multiple Fra-1-bound enhancers showing different molecular and functional features can cooperate to repress gene transcriptionCell & BioscienceJuly 18, 2023Fabienne Bejjani, Emilie Evanno, Samantha Mahfoud, Claire Tolza, Kazem Zibara, Marc Piechaczyk, Isabelle Jariel-Encontreantibodies were from Santa Cruz Biotechnology, as well as control IgG (sc-2025). The anti-p300- (D2X6N, #54062) -CBP (D6C5, #7389) and -CDK7 (#2916) antibodies were from Cell Signaling Technology. The anti-Pol II-PSer5

(5) The Role of PGK1 in Promoting Ischemia/Reperfusion Injury-Induced Microglial M1 Polarization and Inflammation by Regulating GlycolysisNeuroMolecular MedicineJune 1, 2023Wei Cao, Zhengzhe Feng, Deyuan Zhu, Suya Li, Meng Du, Shifei Ye, Dayong Qi, Peng Li, Yan Chen, Yibin Fangagainst PGK1 (CST, 68540S), PKM2 (CST, 4053T), LDHA (CST, 3582T), H3K27ac (CST, 8173T), p300 (CST, 54062S), GAPDH (Proteintech; 60004-1-Ig), and secondary antibodies (Beyotime, A0208). Plasmid Construction

(6) RBM14 as a novel epigenetic-activated tumor oncogene is implicated in the reprogramming of glycolysis in lung cancerWorld Journal of Surgical OncologyApril 14, 2023Yan Hu, Hanshuo Mu, Zhiping Dengprimary antibodies against RBM14 (ab70636, Abcam), YY1 (#63227, Cell Signaling Technology), p300 (#54062, Cell Signaling Technology), and β-actin (ab179467, Abcam) over-night at 4°C. After washing, the secondary

(7) Phosphoglycerate dehydrogenase activates PKM2 to phosphorylate histone H3T11 and attenuate cellular senescenceNature communicationsMarch 10, 2023Wu Y, Tang L, Huang H, Yu Q, Hu B, Wang G, Ge F, Yin T, Li S, Yu X.SA00001-1) were obtained from proteintech; antibodies against PKM2 (1:3000; 4053 S), p300 (1:5000; 54062), and ATF4 (1:1000; 11815) were purchased from Cell Signaling Technology; antibodies against mouse CDKN1A/p21

(8) STRIP2 motivates non-small cell lung cancer progression by modulating the TMBIM6 stability through IGF2BP3 dependentJournal of experimental & clinical cancer research : CRJanuary 13, 2023Zhang X, Chen Q, He Y, Shi Q, Yin C, Xie Y, Yu H, Bao Y, Wang X, Tang C, et al.incubated with specific CBP antibody (cat. no. 7425; Cell Signaling Technology) or P300 antibody (cat. no. 54062; Cell Signaling Technology) to immunoprecipitation chromatin overnight at 4 °C with rotation and followed

(9) Nuclear localization of mitochondrial TCA cycle enzymes modulates pluripotency via histone acetylationNature communicationsDecember 2, 2022Li W, Long Q, Wu H, Zhou Y, Duan L, Yuan H, Ding Y, Huang Y, Wu Y, Huang J, et al.83932,1:100), anti-HA (Cell Signaling Technology, 3724, 1:100), anti-P300 (Cell Signaling Technology, 54062, 1:50), anti-H3K9ac (Cell Signaling Technology, 9649, 1:100), anti-H3K27ac (Cell Signaling Technology,

(10) Artemisitene suppresses rheumatoid arthritis progression via modulating METTL3-mediated N6-methyladenosine modification of ICAM2 mRNA in fibroblast-like synoviocytesClinical and translational medicineDecember 1, 2022Chen J, Lin X, He J, Liu D, He L, Zhang M, Luan H, Hu Y, Tao C, Wang Q.Subsequently, the extracted chromatin was used for fragmentation and immunoprecipitation with p300 (54062, CST, USA) antibody‐conjugated magnetic beads. The eluted DNA was purified and used for further analysis

(11) Bromodomain-containing protein 4 (BRD4) as an epigenetic regulator of fatty acid metabolism genes and ferroptosisCell death & diseaseOctober 29, 2022Yang M, Liu K, Chen P, Zhu H, Wang J, Huang J.(WB) ATP5F1,Proteintech,15999-1-AP,1:500 (WB) H3K27ac,CST,8173,1:50 (IP) Pol II,Abcam,ab264350,1:50 (IP) P300,CST,54062,1:50 (IP) HADH,Abcam,ab110284,1:1000 (WB); 1:50 (IF) ACSL1,Abcam,Ab189939,1:1000 (WB); 1:50 (IF) ACAA1,Abcam,ab1289293,1:1000

(12) EWSR1-ATF1 dependent 3D connectivity regulates oncogenic and differentiation programs in Clear Cell SarcomaNature communicationsApril 27, 2022Möller E, Praz V, Rajendran S, Dong R, Cauderay A, Xing YH, Lee L, Fusco C, Broye LC, Cironi L, et al.Active Motif), -H3K4me3 (#07–473, Millipore), -H3K27ac (#39133, Active motif), -H3K4me1 (ab8895, Abcam), -p300 (#54062, Cell signaling) or -V5 (#13202, Cell signaling) antibodies. Antibody-chromatin complexes were pulled

(13) Early-life undernutrition induces enhancer RNA remodeling in mice liverEpigenetics & ChromatinDecember 1, 2021Yinyu Wang, Yiting Mao, Yiran Zhao, Xianfu Yi, Guolian Ding, Chuanjin Yu, Jianzhong Sheng, Xinmei Liu, Yicong Meng, Hefeng HuangProtein A + G Magnetic beads coupled with anti-p300 antibodies (Cell Signaling Technology, MA, USA, #54062). After reverse crosslinking, ChIP and input DNA fragments were used directly for qPCR with Cry1 (10:85185100–85185350,

(14) An acetylation-enhanced interaction between transcription factor Sox2 and the steroid receptor coactivators facilitates Sox2 transcriptional activity and functionJournal of Biological ChemistryDecember 1, 2021Yuanyong Huang, Xiaoya Duan, Zhen Wang, Yimei Sun, Qingqing Guan, Li Kang, Qiao Zhang, Lan Fang, Jiwen Li, Jiemin Wonganti-SRC-2 antibody (CST, 96687, rabbit), anti-SRC-3 antibody (CST, 2126, rabbit), anti-p300 antibody (CST, 54062, rabbit), anti-GAPDH antibody (Abcam, M20006L, mouse), and anti-Sox2 antibody (Abcam, ab92494, rabbit). WB

(15) Proteomic Screens for Suppressors of Anoikis Identify IL1RAP as a Promising Surface Target in Ewing SarcomaCancer discoveryNovember 1, 2021Zhang HF, Hughes CS, Li W, He JZ, Surdez D, El-Naggar AM, Cheng H, Prudova A, Delaidelli A, Negri GL, et al.ab15289, Abcam), SMARCC1/BAF155 antibody (1:100, #11956, Cell Signaling Technology), p300 antibody (1:50, #54062, CST), H3K27ac antibody (1:100, #39133, Active Motif), or normal rabbit IgG (5 μg/mL, #2729, Cell Signaling Technology

(16) Heterogeneity of subsets in glioblastoma mediated by Smad3 palmitoylationOncogenesisOctober 27, 2021Xiaoqing Fan, Junqi Fan, Haoran Yang, Chenggang Zhao, Wanxiang Niu, Zhiyou Fang, Xueran ChenUSA). ZDHHC19 (orb186631) was sourced from Biorbyt Ltd (Cambridge, UK). Smad3 (9523), E2F4 (40291), EP300 (54062), and β-actin (3700) were purchased from Cell Signaling Technology (Beverly, MA, USA). Flag-tag (F1804)

(17) Proteome-wide profiling of transcriptional machinery on accessible chromatin with biotinylated transposonsScience advancesOctober 22, 2021Zhang H, Qin Z, Yue X, Liu Y, Sun X, Feng J, Xu Z, Zhao J, Li K, Qiu J, et al.antibodies,ChIP-seq antibodies NFKB1,CST,Cas no. 12540 HDAC1,CST,Cas no. D5C6U EP300,CST,Cas no. 54062S RFX1,NOVUS,Cas no. NBP1-52654 MYOD,Thermo Fisher Scientific,Cas no. PA5-34645 ,, Experimental models:

(18) ATF3 promotes the serine synthesis pathway and tumor growth under dietary serine restrictionCell ReportsSeptember 21, 2021Xingyao Li, Daniel Gracilla, Lun Cai, Mingyi Zhang, Xiaolin Yu, Xiaoguang Chen, Junran Zhang, Xiaochun Long, Han-Fei Ding, Chunhong Yan,Cell Signaling,cat#11815; RRID: AB_2616025 p300,Santa Cruz,cat#sc-584; RRID: AB_2293429 p300,Cell Signaling,cat#54062; RRID: AB_2799450 p53,Santa Cruz,cat#sc-126; RRID: AB_628082 PHGDH,Sigma-Aldrich,cat#HPA021241; RRID:

(19) ATF5 and HIF1α cooperatively activate HIF1 signaling pathway in esophageal cancerCell communication and signaling : CCSMay 12, 2021He F, Xiao H, Cai Y, Zhang N.Abcam (Cambridge, MA, USA). Antibodies for PDK1 (13037), PGK1 (68540), CA9 (5649), CBP (7389), P300 (54062), HIF1α (14179), HIF1β (5537) and flag-tag (2368) were procured from Cell Signaling Technology. The

(20) CFTR Cooperative Cis-Regulatory Elements in Intestinal CellsInternational Journal of Molecular SciencesMarch 5, 2021MÃ©gane Collobert, Ozvan Bocher, AnaÃ¯s Le Nabec, Emmanuelle GÃ©nin, Claude FÃ©rec, StÃ©phanie Moisan(Cell Signaling Technology) for 2 h at 4 ◦ C and immunoprecipitation with HNF1α (#89670, Cell Signaling Technology), p300 (#54062, Cell Signaling Technology, Danvers, MA, USA), CDX2 (#12306, Cell Signaling Technology), TCF4 (#2569,

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Nature communications

Cell death & disease

Oncogenesis

Journal of Biological Chemistry

Journal of Biological Chemistry

p300 (D2X6N) Rabbit mAb from Cell Signaling Technology

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Cell Signaling Technology

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p300 (D2X6N) Rabbit mAb from Cell Signaling Technology

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