重组Anti-p53 (phospho S15)抗体[EPR64(N)] - ChIP Grade (ab223868)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR64(N)] to p53 (phospho S15) - ChIP Grade
- Suitable for: ChIP, ICC/IF, IP, Dot blot, WB, ChIC/CUT&RUN-seq
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-p53 (phospho S15)抗体[EPR64(N)] - ChIP Grade
参阅全部 p53 一抗 -
描述
兔单克隆抗体[EPR64(N)] to p53 (phospho S15) - ChIP Grade -
宿主
Rabbit -
经测试应用
适用于: ChIP, ICC/IF, IP, Dot blot, WB, ChIC/CUT&RUN-seqmore details -
种属反应性
与反应: Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: Doxorubicin treated A431 cell lysate. ICC/IF: Doxorubicin treated A431 cells. IP: Doxorubicin treated A431 cell lysate. ChIP: Etoposide treated HCT 116 cells. Dot blot: p53 phospo S15 peptide. ChIC/CUT&RUN-Seq: U2OS cells.
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常规说明
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), 59% PBS -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR64(N) -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab223868于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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ChIP |
Use 5 µg for 25 µg of chromatin.
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ICC/IF |
1/500.
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IP |
1/40.
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Dot blot |
1/1000.
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WB | (1) |
1/5000. Detects a band of approximately 53 kDa (predicted molecular weight: 44 kDa).
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ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
5 µg |
说明 |
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ChIP
Use 5 µg for 25 µg of chromatin. |
ICC/IF
1/500. |
IP
1/40. |
Dot blot
1/1000. |
WB
1/5000. Detects a band of approximately 53 kDa (predicted molecular weight: 44 kDa). |
ChIC/CUT&RUN-seq
Use at an assay dependent concentration. 5 µg |
靶标
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功能
Acts as a tumor suppressor in many tumor types; induces growth arrest or apoptosis depending on the physiological circumstances and cell type. Involved in cell cycle regulation as a trans-activator that acts to negatively regulate cell division by controlling a set of genes required for this process. One of the activated genes is an inhibitor of cyclin-dependent kinases. Apoptosis induction seems to be mediated either by stimulation of BAX and FAS antigen expression, or by repression of Bcl-2 expression. Implicated in Notch signaling cross-over. Isoform 2 enhances the transactivation activity of isoform 1 from some but not all TP53-inducible promoters. Isoform 4 suppresses transactivation activity and impairs growth suppression mediated by isoform 1. Isoform 7 inhibits isoform 1-mediated apoptosis. -
组织特异性
Ubiquitous. Isoforms are expressed in a wide range of normal tissues but in a tissue-dependent manner. Isoform 2 is expressed in most normal tissues but is not detected in brain, lung, prostate, muscle, fetal brain, spinal cord and fetal liver. Isoform 3 is expressed in most normal tissues but is not detected in lung, spleen, testis, fetal brain, spinal cord and fetal liver. Isoform 7 is expressed in most normal tissues but is not detected in prostate, uterus, skeletal muscle and breast. Isoform 8 is detected only in colon, bone marrow, testis, fetal brain and intestine. Isoform 9 is expressed in most normal tissues but is not detected in brain, heart, lung, fetal liver, salivary gland, breast or intestine. -
疾病相关
Note=TP53 is found in increased amounts in a wide variety of transformed cells. TP53 is frequently mutated or inactivated in about 60% of cancers. TP53 defects are found in Barrett metaplasia a condition in which the normally stratified squamous epithelium of the lower esophagus is replaced by a metaplastic columnar epithelium. The condition develops as a complication in approximately 10% of patients with chronic gastroesophageal reflux disease and predisposes to the development of esophageal adenocarcinoma.
Defects in TP53 are a cause of esophageal cancer (ESCR) [MIM:133239].
Defects in TP53 are a cause of Li-Fraumeni syndrome (LFS) [MIM:151623]. LFS is an autosomal dominant familial cancer syndrome that in its classic form is defined by the existence of a proband affected by a sarcoma before 45 years with a first degree relative affected by any tumor before 45 years and another first degree relative with any tumor before 45 years or a sarcoma at any age. Other clinical definitions for LFS have been proposed (PubMed:8118819 and PubMed:8718514) and called Li-Fraumeni like syndrome (LFL). In these families affected relatives develop a diverse set of malignancies at unusually early ages. Four types of cancers account for 80% of tumors occurring in TP53 germline mutation carriers: breast cancers, soft tissue and bone sarcomas, brain tumors (astrocytomas) and adrenocortical carcinomas. Less frequent tumors include choroid plexus carcinoma or papilloma before the age of 15, rhabdomyosarcoma before the age of 5, leukemia, Wilms tumor, malignant phyllodes tumor, colorectal and gastric cancers.
Defects in TP53 are involved in head and neck squamous cell carcinomas (HNSCC) [MIM:275355]; also known as squamous cell carcinoma of the head and neck.
Defects in TP53 are a cause of lung cancer (LNCR) [MIM:211980].
Defects in TP53 are a cause of choroid plexus papilloma (CPLPA) [MIM:260500]. Choroid plexus papilloma is a slow-growing benign tumor of the choroid plexus that often invades the leptomeninges. In children it is usually in a lateral ventricle but in adults it is more often in the fourth ventricle. Hydrocephalus is common, either from obstruction or from tumor secretion of cerebrospinal fluid. If it undergoes malignant transformation it is called a choroid plexus carcinoma. Primary choroid plexus tumors are rare and usually occur in early childhood.
Defects in TP53 are a cause of adrenocortical carcinoma (ADCC) [MIM:202300]. ADCC is a rare childhood tumor of the adrenal cortex. It occurs with increased frequency in patients with the Beckwith-Wiedemann syndrome and is a component tumor in Li-Fraumeni syndrome. -
序列相似性
Belongs to the p53 family. -
结构域
The nuclear export signal acts as a transcriptional repression domain. The TADI and TADII motifs (residues 17 to 25 and 48 to 56) correspond both to 9aaTAD motifs which are transactivation domains present in a large number of yeast and animal transcription factors. -
翻译后修饰
Acetylated. Acetylation of Lys-382 by CREBBP enhances transcriptional activity. Deacetylation of Lys-382 by SIRT1 impairs its ability to induce proapoptotic program and modulate cell senescence.
Phosphorylation on Ser residues mediates transcriptional activation. Phosphorylated by HIPK1 (By similarity). Phosphorylation at Ser-9 by HIPK4 increases repression activity on BIRC5 promoter. Phosphorylated on Thr-18 by VRK1. Phosphorylated on Ser-20 by CHEK2 in response to DNA damage, which prevents ubiquitination by MDM2. Phosphorylated on Thr-55 by TAF1, which promotes MDM2-mediated degradation. Phosphorylated on Ser-46 by HIPK2 upon UV irradiation. Phosphorylation on Ser-46 is required for acetylation by CREBBP. Phosphorylated on Ser-392 following UV but not gamma irradiation. Phosphorylated upon DNA damage, probably by ATM or ATR. Phosphorylated on Ser-15 upon ultraviolet irradiation; which is enhanced by interaction with BANP.
Dephosphorylated by PP2A-PPP2R5C holoenzyme at Thr-55. SV40 small T antigen inhibits the dephosphorylation by the AC form of PP2A.
May be O-glycosylated in the C-terminal basic region. Studied in EB-1 cell line.
Ubiquitinated by MDM2 and SYVN1, which leads to proteasomal degradation. Ubiquitinated by RFWD3, which works in cooperation with MDM2 and may catalyze the formation of short polyubiquitin chains on p53/TP53 that are not targeted to the proteasome. Ubiquitinated by MKRN1 at Lys-291 and Lys-292, which leads to proteasomal degradation. Deubiquitinated by USP10, leading to its stabilization. Ubiquitinated by TRIM24, which leads to proteasomal degradation. Ubiquitination by TOPORS induces degradation. Deubiquitination by USP7, leading to stabilization. Isoform 4 is monoubiquitinated in an MDM2-independent manner.
Monomethylated at Lys-372 by SETD7, leading to stabilization and increased transcriptional activation. Monomethylated at Lys-370 by SMYD2, leading to decreased DNA-binding activity and subsequent transcriptional regulation activity. Lys-372 monomethylation prevents interaction with SMYD2 and subsequent monomethylation at Lys-370. Dimethylated at Lys-373 by EHMT1 and EHMT2. Monomethylated at Lys-382 by SETD8, promoting interaction with L3MBTL1 and leading to repress transcriptional activity. Demethylation of dimethylated Lys-370 by KDM1A prevents interaction with TP53BP1 and represses TP53-mediated transcriptional activation.
Sumoylated by SUMO1. -
细胞定位
Cytoplasm; Cytoplasm. Nucleus. Nucleus > PML body. Endoplasmic reticulum. Interaction with BANP promotes nuclear localization. Recruited into PML bodies together with CHEK2; Nucleus. Cytoplasm. Localized in both nucleus and cytoplasm in most cells. In some cells, forms foci in the nucleus that are different from nucleoli; Nucleus. Cytoplasm. Localized in the nucleus in most cells but found in the cytoplasm in some cells; Nucleus. Cytoplasm. Localized mainly in the nucleus with minor staining in the cytoplasm; Nucleus. Cytoplasm. Predominantly nuclear but localizes to the cytoplasm when expressed with isoform 4 and Nucleus. Cytoplasm. Predominantly nuclear but translocates to the cytoplasm following cell stress. - Information by UniProt
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数据库链接
- Entrez Gene: 7157 Human
- Omim: 191170 Human
- SwissProt: P04637 Human
- Unigene: 654481 Human
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别名
- Antigen NY-CO-13 antibody
- BCC7 antibody
- Cellular tumor antigen p53 antibody
see all
图片
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ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 U2OS cells treated with Etoposide (5μM 18h) and 5 µg of ab223868 [EPR64(N]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
Additional screenshots of mapped reads can be downloaded here.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
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All lanes : Anti-p53 (phospho S15) antibody [EPR64(N)] - ChIP Grade (ab223868) at 1/5000 dilution
Lane 1 : Untreated A431 (human epidermoid carcinoma cell line), whole cell lysate
Lane 2 : A431 (human epidermoid carcinoma cell line) treated with 1 µg/ml doxorubicin for 24 hours, whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 44 kDa
Observed band size: 53 kDa why is the actual band size different from the predicted?
Exposure time: 1 secondBlocking/Dilution buffer: 2% BSA/TBST.
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Immunofluorescent analysis of 4% PFA-fixed A431 (human epidermoid carcinoma cell line) cells labeling p53 (phospho S15) with ab223868 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing increased nuclear staining on A431 cells treated with 1 μg/ml doxorubicin for 24 hours.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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Dot blot analysis of p53 (phospho S15) labeled with ab223868 at 1/1000 dilution.
Lane 1: p53 (phospho S15) peptide;
Lane 2: p53 non-phospho peptide;
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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p53 (phospho S15) was immunoprecipitated from 0.35 mg of A431 (human epidermoid carcinoma cell line) whole cell lysate with ab223868 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab223868 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: A431 treated with 1 μg/ml doxorubicin for 24 hours, whole cell lysate 10 µg (Input).
Lane 2: ab223868 IP in A431 treated with 1 μg/ml doxorubicin for 24 hours, whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab223868 in A431 treated with 1 μg/ml doxorubicin for 24 hours, whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
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Chromatin was prepared from HCT 116 (human colorectal carcinoma cell line) cells untreated or treated with 50 µM Etoposide for 6 hours according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. ChIP was performed with 25 mg of chromatin, 5 µg of ab223868 anti-p53 (phospho S15) (blue), and 20 µl of A/G sepharose beads slurry (10 µl of sepharose A beads + 10 µl of sepharose G beads). Rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (SYBR approach).
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (6)
ab223868 被引用在 6 文献中.
- Wu WY et al. SSBP1 drives high fructose-induced glomerular podocyte ferroptosis via activating DNA-PK/p53 pathway. Redox Biol 52:102303 (2022). PubMed: 35390676
- Wang S et al. Fraxinellone Induces Hepatotoxicity in Zebrafish through Oxidative Stress and the Transporters Pathway. Molecules 27:N/A (2022). PubMed: 35566003
- Zhang D et al. Runt-related transcription factor 1 (Runx1) aggravates pathological cardiac hypertrophy by promoting p53 expression. J Cell Mol Med 25:7867-7877 (2021). PubMed: 34190420
- He H et al. Copper Oxide Nanoparticles Induce Oxidative DNA Damage and Cell Death via Copper Ion-Mediated P38 MAPK Activation in Vascular Endothelial Cells. Int J Nanomedicine 15:3291-3302 (2020). PubMed: 32494130
- Sola M et al. Tau affects P53 function and cell fate during the DNA damage response. Commun Biol 3:245 (2020). PubMed: 32427887
- Jin J et al. Oxidative stress induces p38MAPK-dependent senescence in the feto-maternal interface cells. Placenta 67:15-23 (2018). PubMed: 29941169