Anti-KAP1 抗体 [20C1] - ChIP Grade (ab22553)

製品の概要

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab22553 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ChIP Use at an assay dependent dilution.
IHC-P Use a concentration of 5 µg/ml.
ICC/IF Use a concentration of 2 - 4 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 88.5 kDa).
EMSA Use at an assay dependent dilution.
IP Use at an assay dependent concentration.

ターゲット情報

  • 機能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.
  • 組織特異性Expressed in all tissues tested including spleen, thymus, prostate, testis, ovary, small intestine, colon and peripheral blood leukocytes.
  • パスウェイProtein modification; protein sumoylation.
  • 配列類似性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.
  • ドメイン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.
  • 翻訳後修飾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.
  • 細胞内局在Nucleus. Associated with centromeric heterochromatin during cell differentiation through CBX1.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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

Anti-KAP1 antibody [20C1] - ChIP Grade 画像

  • Anti-KAP1 antibody [20C1] - ChIP Grade (ab22553) at 1/500 dilution + HeLa whole cell lysate

    Secondary
    Goat anti-mouse-HRP at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 88.5 kDa


    Exposure time : 1 second

    This image is courtesy of an anonymous Abreview

    See Abreview

  • Immunofluorescence analysis of A549 cells, staining KAP1 (green) with ab22553.
    Cells were fixed in 4% paraformaldehyde followed by permeabilization in 0.1% Triton X-100/PBS. Cells were incubated in primary antibody and a FITC-conjugated goat anti-mouse secondary antibody was used to detect staining. Nuclei were stained with DAPI (blue).

  • IHC image of KAP1 staining in Human normal spleen formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab22553, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Anti-KAP1 antibody [20C1] - ChIP Grade (ab22553) 使用論文

This product has been referenced in:
  • Thompson PJ  et al. hnRNP K Coordinates Transcriptional Silencing by SETDB1 in Embryonic Stem Cells. PLoS Genet 11:e1004933 (2015). WB, IP, ChIP . Read more (PubMed: 25611934) »
  • Gjyshi O  et al. Activated Nrf2 Interacts with Kaposi's Sarcoma-Associated Herpesvirus Latency Protein LANA-1 and Host Protein KAP1 To Mediate Global Lytic Gene Repression. J Virol 89:7874-92 (2015). Read more (PubMed: 25995248) »

See all 19 Publications for this product

Product Wall

Application Western blot
Sample Human Cell lysate - whole cell (B Cells)
Gel Running Conditions Reduced Denaturing (10% SDS-PAGE)
Loading amount 50 µg
Specification B Cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Oct 26 2016


Unfortunately, the epitope for this ab22553Anti-KAP1[20C1] monoclonal antibody has not been specifically mapped.       

The immunogen used is a recombinant fragment, corresponding to amino acids 60-383 of Human KAP1. This was from isoform...

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Thank you for contacting us. No, these antibodies have not been tested with overexpression or knockdown cells.


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Thank you for contacting us and reporting the problems the customer has been experiencing with the Anti-KAP1 antibody [20C1] - ChIP Grade (ab22553).

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HeLa)
Loading amount 50 µg
Specification HeLa
Gel Running Conditions Reduced Denaturing (12%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

投稿 Apr 16 2012

Thank you for submitting your Abreview regarding your experience with ab22553. I'm sorry the ChIP experiment did not work as expected. It appears the vial of ab22553 you received was not working as expected so if you submitted your review within 6 mont...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"