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Epigenetics and Nuclear Signaling Chromatin Binding Proteins Methylated DNA

Anti-KAP1抗体(ab3831)

  • Datasheet
Reviews (4)Q&A (4)References (8)

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Western blot - Anti-KAP1 antibody (ab3831)
  • Immunocytochemistry/ Immunofluorescence - Anti-KAP1 antibody (ab3831)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAP1 antibody (ab3831)

Key features and details

  • Goat polyclonal to KAP1
  • Suitable for: ICC/IF, IHC-P, WB
  • Reacts with: Mouse, Human
  • Isotype: IgG

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概述

  • 产品名称

    Anti-KAP1抗体
    参阅全部 KAP1 一抗
  • 描述

    山羊多克隆抗体to KAP1
  • 宿主

    Goat
  • 经测试应用

    适用于: ICC/IF, IHC-P, WBmore details
  • 种属反应性

    与反应: Mouse, Human
  • 免疫原

    Synthetic peptide: SSQELSGGPGDGP, corresponding to amino acids 741-753 of KAP 1.

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • 阳性对照

    • 购买相配的WB阳性对照Recombinant Human KAP1 protein
    • WB: HepG2 lysate. IHC-P: Human breast tissue.
  • 常规说明

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

性能

  • 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 or -80°C. Avoid freeze / thaw cycle.
  • 存储溶液

    pH: 7.30
    Preservative: 0.02% Sodium azide
    Constituents: Tris buffered saline, 0.5% BSA
  • Concentration information loading...
  • 纯度

    Immunogen affinity purified
  • 纯化说明

    Purified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide.
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Epigenetics and Nuclear Signaling
    • Chromatin Binding Proteins
    • Methylated DNA
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteasome / Ubiquitin
    • Ubiquitin E3 Enzymes
    • RING Finger E3 Ligase
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Transcription Factors
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Other
    • Cancer
    • Cell Death
    • Apoptosis
    • Other
    • Epigenetics and Nuclear Signaling
    • Bromodomains

相关产品

  • Compatible Secondaries

    • Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (ab150129)
    • Donkey Anti-Goat IgG H&L (HRP) (ab205723)
  • Isotype control

    • Goat IgG, polyclonal - Isotype Control (ab37373)
  • Recombinant Protein

    • Recombinant Human KAP1 protein (ab131899)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab3831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
ICC/IF (1)
Use a concentration of 5 µg/ml.
IHC-P
Use a concentration of 4 - 6 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB (2)
Use a concentration of 1 - 3 µg/ml. Detects a band of approximately 110 kDa (predicted molecular weight: 100 kDa).

1 hour primary incubation is recommended for this product.

说明
ICC/IF
Use a concentration of 5 µg/ml.
IHC-P
Use a concentration of 4 - 6 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB
Use a concentration of 1 - 3 µg/ml. Detects a band of approximately 110 kDa (predicted molecular weight: 100 kDa).

1 hour primary incubation is recommended for this product.

靶标

  • 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.
  • Target information above from: UniProt accession Q13263 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 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
    • 别名

      • E3 SUMO protein ligase TRIM28 antibody
      • E3 SUMO-protein ligase TRIM28 antibody
      • FLJ29029 antibody
      • KAP 1 antibody
      • KAP-1 antibody
      • KRAB associated protein 1 antibody
      • KRAB interacting protein 1 antibody
      • KRAB-associated protein 1 antibody
      • KRAB-interacting protein 1 antibody
      • KRIP 1 antibody
      • KRIP-1 antibody
      • KRIP1 antibody
      • Nuclear corepressor KAP 1 antibody
      • Nuclear corepressor KAP-1 antibody
      • RING finger protein 96 antibody
      • RNF96 antibody
      • TF1B antibody
      • TIF1 beta antibody
      • TIF1-beta antibody
      • TIF1B antibody
      • TIF1B_HUMAN antibody
      • Transcription intermediary factor 1 beta antibody
      • Transcription intermediary factor 1-beta antibody
      • Trim28 antibody
      • Tripartite motif containing 28 antibody
      • tripartite motif containing protein 28 antibody
      • Tripartite motif-containing protein 28 antibody
      see all

    图片

    • Western blot - Anti-KAP1 antibody (ab3831)
      Western blot - Anti-KAP1 antibody (ab3831)
      ab3831 staining (0.5µg/ml) of HepG2 lysate (RIPA buffer, 30µg total protein per lane).  Primary incubated for 1 hour.  Detected by western blot using chemiluminescence. ab3831 staining (0.5µg/ml) of HepG2 lysate (RIPA buffer, 30µg total protein per lane). Primary incubated for 1 hour. Detected by western blot using chemiluminescence.
    • Immunocytochemistry/ Immunofluorescence - Anti-KAP1 antibody (ab3831)
      Immunocytochemistry/ Immunofluorescence - Anti-KAP1 antibody (ab3831)
      TIF1b is uniformly distributed in the two pronuclei of the mouse embryo at the late zygote stage. Mouse zygotes were processed for immunostaining using the KAP1 antibody. DNA is shown in blue. The dilution used was 1 to 100 in PBS-Tween with 3% BSA. This is part of the review submitted by ME Torres-Padilla on 27 July 2004.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAP1 antibody (ab3831)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAP1 antibody (ab3831)

      Paraffin embedded human breast tissue stained for KAP1 using ab3831 at 2 µg/ml in immunohistochemical analysis. Steamed antigen retrieval with Tris/EDTA buffer pH 9, HRP-staining.

    实验方案

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    数据表及文件

    • Datasheet download

      Download

    文献 (8)

    发表研究结果有使用 ab3831?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab3831 被引用在 8 文献中.

    • Himeda CL  et al. Targeted epigenetic repression by CRISPR/dSaCas9 suppresses pathogenic DUX4-fl expression in FSHD. Mol Ther Methods Clin Dev 20:298-311 (2021). PubMed: 33511244
    • Himeda CL  et al. CRISPR/dCas9-mediated Transcriptional Inhibition Ameliorates the Epigenetic Dysregulation at D4Z4 and Represses DUX4-fl in FSH Muscular Dystrophy. Mol Ther 24:527-35 (2016). ChIP ; Human . PubMed: 26527377
    • Nakajima NI  et al. Pre-exposure to ionizing radiation stimulates DNA double strand break end resection, promoting the use of homologous recombination repair. PLoS One 10:e0122582 (2015). WB . PubMed: 25826455
    • Yüce Ö & West SC Senataxin, defective in the neurodegenerative disorder ataxia with oculomotor apraxia 2, lies at the interface of transcription and the DNA damage response. Mol Cell Biol 33:406-17 (2013). WB . PubMed: 23149945
    • Maruyama A  et al. The novel Nrf2-interacting factor KAP1 regulates susceptibility to oxidative stress by promoting the Nrf2-mediated cytoprotective response. Biochem J 436:387-97 (2011). PubMed: 21382013
    • Noon AT  et al. 53BP1-dependent robust localized KAP-1 phosphorylation is essential for heterochromatic DNA double-strand break repair. Nat Cell Biol 12:177-84 (2010). PubMed: 20081839
    • Frietze S  et al. ZNF274 recruits the histone methyltransferase SETDB1 to the 3' ends of ZNF genes. PLoS One 5:e15082 (2010). ChIP-sequencing ; Human . PubMed: 21170338
    • Torres-Padilla ME & Zernicka-Goetz M Role of TIF1alpha as a modulator of embryonic transcription in the mouse zygote. J Cell Biol 174:329-38 (2006). WB, IHC-FrFl, IP ; Mouse . PubMed: 16880268

    客户评价及客户问答

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    1-8 of 8 Abreviews or Q&A

    Western blot abreview for Anti-KAP1 antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (MEF)
    Gel Running Conditions
    Reduced Denaturing (4-20%)
    Loading amount
    10 µg
    Specification
    MEF
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    提交于 Apr 24 2019

    Immunocytochemistry/ Immunofluorescence abreview for Anti-KAP1 antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Permeabilization
    Yes - 0.5% Triton 5 minutes
    Specification
    HeLa
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    提交于 Oct 26 2017

    Western blot abreview for Anti-KAP1 antibody - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK293)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    10 µg
    Treatment
    1uM CPT 1hr
    Specification
    HEK293
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    提交于 Jul 05 2016

    ChIP abreview for Anti-KAP1 antibody - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    ChIP
    Sample
    Human Cell lysate - nuclear (K562 cells)
    Specification
    K562 cells
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Specification of the cross-linking agent: formaldehyde
    Detection step
    Other
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Dr. Seth Frietze

    Verified customer

    提交于 Mar 08 2011

    Question

    Thank you for kind advises about #ab3831. I have informed your comments to our customer. You recommend following modifications on his protocol. To decrease the background, try blocking with BSA rather than milk. I also suggest decreasing the concentrations of both the primary and secondary antibodies and decrease the incubation periods. In addition, increase the number of washes. They have already changed these condition, however these modification did not improve his results. Additionally, you also suggested running a positive control, and HepG2 whole cell lysate is recommended as positive control. But, please tell us which is a positive control of the KAP1 antibody in mouse extract, because they use a extract from mouse cell line NIH3T3. And please tell us which samples from mouse were used in your test if you possible. I'd like to know whether #ab3831 certainly work on mouse sample. Please let me know. Thank you.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 25 2005

    Answer

    Thank you for your email. This antibody was originally characterized for application in Western blotting using human samples (HepG2 whole cell lysate). We received feedback from a customer who successfully used ab3831 with mouse samples (mouse embryos) in Western blotting and IF. You may read this review by clicking on the reviews tab located on the online datasheet. You may also send an email to the reviewer by clicking on the name. As I previously mentioned, I would suggest using HepG2 whole cell lysate as a positive control. On the online datasheet for ab3831, there is also a nice Western blot image showing the results with this lysate. Please let me know if you need further assistance.

    Read More

    Abcam Scientific Support

    回复于 Jan 28 2005

    Question

    We received a compliant for #AB3831 from our customer who bought it in this month. The Lot#s was #87823. They demonstrated the Western Blotting Assay using this antibody as primary antibody. Our customer reported that signals were not observed at all. Their experimental conditions was as follows. Mouse NIH3T3 cell line was used as sample. They prepared the sample as crude whole cell lysate. 10-20ug of protein was included in their sample. Sample ware not induced in their expression. Membrane was blocked by 5% skim milk for one hour at room temperature. About primary antibody Dilution ratio was 1:2000. Reaction was performed for over night at room temperature. About secondary antibody; HRP-alfa-goat IgG, F(ab')2 fragment specific (Jackson) Secondary antibody was diluted up to 10000 fold. They could get good results in other experiment using this secondary antibody. Background signal was high and they also detect non-specific band, however they could observe only week band of their sample. Their sample was stained by other primary antibodies ( Anti-BAF155(R-18), goat poly Ab, Santa Cruz; Anti NUP50, goat poly Ab, Abcam ) in same method , however they didn't use positive and negative control. And they hope that it is replaced with other product (#ab10483) if possible. Could you send another product as replacement? Please let me know your concern. Thank you.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 20 2005

    Answer

    Thank you for your email and I'm sorry to hear that your customer is experiencing difficulty with ab3831. At this point I would like to make some suggestions to try to assist your customer. To decrease the background, try blocking with BSA rather than milk. I also suggest decreasing the concentrations of both the primary and secondary antibodies and decrease the incubation periods. In addition, increase the number of washes. I also strongly suggest running a positive control. Ab3831 was characterized using HepG2 whole cell lysate, which is recommended as positive control. If these suggestions do not help, please let me know.

    Read More

    Abcam Scientific Support

    回复于 Jan 20 2005

    Question

    I would like to ask for some technical information about your KAP-1 antibody (goat polyclonal, cat no. ab3831). From western blot it seems KAP-1 migrates as a 120kDa band. However its molecular mass should be 88.5kDa. Do you have the reason why it shifts differently from its calculated mass? Thanks.

    Read More

    Abcam community

    Verified customer

    Asked on Oct 04 2004

    Answer

    Thank you for your enquiry. According to Swiss Prot and Ugene the predicted molecular weight of this target should be approximately 100 kDa. We have updated the relevant public facing site in order to reflect this data. However, we would like to emphasize that this antibody has been tested and characterized on HepG2 (human hepatocellular cancer cell line). According to Swiss-Prot database, there are alternative splice variants and isoforms which may cause shift in the band size. We would like to thank you for bringing this matter to our attention.

    Read More

    Abcam Scientific Support

    回复于 Oct 05 2004

    Question

    I would like to know if this antibody cross reacts with TIF 1 alpha please?

    Read More

    Abcam community

    Verified customer

    Asked on Oct 01 2003

    Answer

    By BLAST analysis I can find no significant similarity between the epitope used to generate this antibody (Peptide with sequence SSQELSGGPGDGP, from C Terminus of the protein sequence according) and any epitopes in TIF 1 alpha. I therfore think it is highly unlikely that the antibody cross reacts with TIF 1 alpha.

    Read More

    Abcam Scientific Support

    回复于 Oct 01 2003

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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