重组Anti-KAP1 (phospho S824)抗体[EPR5248] (ab133440)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5248] to KAP1 (phospho S824)
- Suitable for: WB, IP, Dot blot
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
概述
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产品名称
Anti-KAP1 (phospho S824)抗体[EPR5248]
参阅全部 KAP1 一抗 -
描述
兔单克隆抗体[EPR5248] to KAP1 (phospho S824) -
宿主
Rabbit -
经测试应用
适用于: WB, IP, Dot blotmore details -
种属反应性
与反应: Mouse, 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: WT HAP1 cell lysate (+/- Bleomycin); HeLa cell lysate (+/- Bleomycin). Dot blot: KAP1 (phospho S824) phospho peptide. IP: HeLa treated with 3uM etoposide for 1h whole cell lysate.
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常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
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 -20°C. Stable for 12 months at -20°C. -
存储溶液
pH: 7.20
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
EPR5248 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Isotype control
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab133440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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WB |
1/1000. Detects a band of approximately 110 kDa (predicted molecular weight: 88 kDa).
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IP |
1/10 - 1/100.
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Dot blot |
Use at an assay dependent concentration.
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说明 |
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WB
1/1000. Detects a band of approximately 110 kDa (predicted molecular weight: 88 kDa). |
IP
1/10 - 1/100. |
Dot blot
Use at an assay dependent concentration. |
靶标
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功能
Nuclear corepressor for KRAB domain-containing zinc finger proteins (KRAB-ZFPs). Mediates gene silencing by recruiting CHD3, a subunit of the nucleosome remodeling and deacetylation (NuRD) complex, and SETDB1 (which specifically methylates histone H3 at 'Lys-9' (H3K9me)) to the promoter regions of KRAB target genes. Enhances transcriptional repression by coordinating the increase in H3K9me, the decrease in histone H3 'Lys-9 and 'Lys-14' acetylation (H3K9ac and H3K14ac, respectively) and the disposition of HP1 proteins to silence gene expression. Recruitment of SETDB1 induces heterochromatinization. May play a role as a coactivator for CEBPB and NR3C1 in the transcriptional activation of ORM1. Also corepressor for ERBB4. Inhibits E2F1 activity by stimulating E2F1-HDAC1 complex formation and inhibiting E2F1 acetylation. May serve as a partial backup to prevent E2F1-mediated apoptosis in the absence of RB1. Important regulator of CDKN1A/p21(CIP1). Has E3 SUMO-protein ligase activity toward itself via its PHD-type zinc finger. -
组织特异性
Expressed in all tissues tested including spleen, thymus, prostate, testis, ovary, small intestine, colon and peripheral blood leukocytes. -
通路
Protein modification; protein sumoylation. -
序列相似性
Belongs to the TRIM/RBCC family.
Contains 2 B box-type zinc fingers.
Contains 1 bromo domain.
Contains 1 PHD-type zinc finger.
Contains 1 RING-type zinc finger. -
结构域
The HP1 box is both necessary and sufficient for HP1 binding.
The PHD-type zinc finger enhances CEBPB transcriptional activity. The PHD-type zinc finger, the HP1 box and the bromo domain, function together to assemble the machinery required for repression of KRAB domain-containing proteins. Acts as an intramolecular SUMO E3 ligase for autosumoylation of bromodomain.
The RING-finger-B Box-coiled-coil/tripartite motif (RBCC/TRIM motif) is required for interaction with the KRAB domain of KRAB-zinc finger proteins. Binds four zinc ions per molecule. The RING finger and the N-terminal of the leucine zipper alpha helical coiled-coil region of RBCC are required for oligomerization.
Contains one Pro-Xaa-Val-Xaa-Leu (PxVxL) motif, which is required for interaction with chromoshadow domains. This motif requires additional residues -7, -6, +4 and +5 of the central Val which contact the chromoshadow domain. -
翻译后修饰
Phosphorylated upon DNA damage, probably by ATM or ATR. ATM-induced phosphorylation on Ser-824 represses sumoylation leading to the de-repression of expression of a subset of genes involved in cell cycle control and apoptosis in response to genotoxic stress. Dephosphorylation by the phosphatases, PPP1CA and PP1CB forms, allows sumoylation and expression of TRIM28 target genes.
Sumoylation/desumoylation events regulate TRIM28-mediated transcriptional repression. Sumoylation is required for interaction with CHD3 and SETDB1 and the corepressor activity. Represses and is repressed by Ser-824 phosphorylation. Enhances the TRIM28 corepressor activity, inhibiting transcriptional activity of a number of genes including GADD45A and CDKN1A/p21. Lys-554, Lys-779 and Lys-804 are the major sites of sumoylation. In response to Dox-induced DNA damage, enhanced phosphorylation on Ser-824 prevents sumoylation and allows de-repression of CDKN1A/p21. -
细胞定位
Nucleus. Associated with centromeric heterochromatin during cell differentiation through CBX1. - Information by UniProt
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数据库链接
- Entrez Gene: 10155 Human
- Entrez Gene: 21849 Mouse
- Omim: 601742 Human
- SwissProt: Q13263 Human
- SwissProt: Q62318 Mouse
- Unigene: 467408 Human
- Unigene: 15701 Mouse
- Unigene: 398345 Mouse
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别名
- E3 SUMO protein ligase TRIM28 antibody
- E3 SUMO-protein ligase TRIM28 antibody
- FLJ29029 antibody
see all
图片
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All lanes : Anti-KAP1 (phospho S824) antibody [EPR5248] (ab133440) at 1/1000 dilution
Lane 1 : Untreated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HeLa treated with 5µM etoposide for 8 hours whole cell lysate
Lane 3 : HeLa treated with 5µM etoposide for 8 hours whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 88 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?
Exposure time: 7 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST.
ab181602 was used as a GAPDH loading control.
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All lanes : Anti-KAP1 (phospho S824) antibody [EPR5248] (ab133440) at 1/1000 dilution (Purified)
Lane 1 : Untreated NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) treated with 3µM etoposide for 1 hour whole cell lysate
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) treated with 3µM etoposide for 1 hour whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 88 kDa -
Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: Wild type HAP1 + DMSO whole cell lysate (20 µg)
Lane 3: Wild type HAP1 + Bleomycin whole cell lysate (20 µg)
Lane 4: KAP1 knockout HAP1 whole cell lysate (20 µg)
Lane 5: KAP1 knockout HAP1 + DMSO whole cell lysate (20 µg)
Lane 6: KAP1 knockout HAP1 + Bleomycin whole cell lysate (20 µg)
Lane 7: HeLa + DMSO whole cell lysate (20 µg)
Lane 8: HeLa + Bleomycin whole cell lysate (20 µg)Lanes 1 - 8: Merged signal (red and green). Green - ab133440 observed at 105 kDa. Red - loading control, ab130007, observed at 125 kDa.
ab133440 was shown to specifically react with KAP1 in wild type cells as signal was lost in KAP1 knockout cells. Wild-type and KAP1 knockout samples were subjected to SDS-PAGE. ab133440 and ab130007 (Mouse anti-vinculin loading control) were incubated overnight at 4°C both at a 1/20000 dilution. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging. Treated with 30 µg/mL Bleomycin in DMSO for 30 minutes.
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Purified ab133440 at 1/50 dilution (2µg) immunoprecipitating KAP1 in HeLa treated with 3uM etoposide for 1h whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) treated with 3uM etoposide for 1h whole cell lysate 10µg
Lane 2 (+): ab133440 + HeLa treated with 3uM etoposide for 1h whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32132 in HeLa treated with 3uM etoposide for 1h whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 110 kDa -
Dot blot analysis of KAP1 (phospho S824) phospho peptide (Lane 1) and KAP1 non-phospho peptide (Lane 2) labelling KAP1 (phospho S824) with Unpurified ab133440 at a dilution of 1/1000. A Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1/20,000. Blocking buffer: 5% NFDM/TBST. Dilution buffer: 5% NFDM /TBST.
数据表及文件
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SDS download
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Datasheet download
文献 (10)
ab133440 被引用在 10 文献中.
- Migliozzi S et al. Integrative multi-omics networks identify PKCδ and DNA-PK as master kinases of glioblastoma subtypes and guide targeted cancer therapy. Nat Cancer 4:181-202 (2023). PubMed: 36732634
- Roulston A et al. RP-3500: A Novel, Potent, and Selective ATR Inhibitor that is Effective in Preclinical Models as a Monotherapy and in Combination with PARP Inhibitors. Mol Cancer Ther 21:245-256 (2022). PubMed: 34911817
- Zimmermann A et al. A New Class of Selective ATM Inhibitors as Combination Partners of DNA Double-Strand Break Inducing Cancer Therapies. Mol Cancer Ther 21:859-870 (2022). PubMed: 35405736
- Fraser CR et al. Radiofluorination of a highly potent ATM inhibitor as a potential PET imaging agent. EJNMMI Res 12:50 (2022). PubMed: 35962885
- Haines E et al. DNA-PK inhibitor peposertib enhances p53-dependent cytotoxicity of DNA double-strand break inducing therapy in acute leukemia. Sci Rep 11:12148 (2021). PubMed: 34108527
- Dunlop CR et al. Complete loss of ATM function augments replication catastrophe induced by ATR inhibition and gemcitabine in pancreatic cancer models. Br J Cancer 123:1424-1436 (2020). PubMed: 32741974
- Frohns F et al. Differences in the Response to DNA Double-Strand Breaks between Rod Photoreceptors of Rodents, Pigs, and Humans. Cells 9:N/A (2020). PubMed: 32290532
- Sun Q et al. Therapeutic Implications of p53 Status on Cancer Cell Fate Following Exposure to Ionizing Radiation and the DNA-PK Inhibitor M3814. Mol Cancer Res 17:2457-2468 (2019). PubMed: 31551253
- Ensminger M et al. DNA breaks and chromosomal aberrations arise when replication meets base excision repair. J Cell Biol 206:29-43 (2014). PubMed: 24982429
- Geuting V et al. ATM release at resected double-strand breaks provides heterochromatin reconstitution to facilitate homologous recombination. PLoS Genet 9:e1003667 (2013). WB ; Human . PubMed: 23935532