製品の概要

  • 製品名Anti-KAP1 antibody - ChIP Grade
    KAP1 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to KAP1 - ChIP Grade
  • アプリケーション適用あり: WB, IP, IHC-P, ICC, ICC/IF, ChIP, ChIP/Chipmore details
  • 種交差性
    交差種: Mouse, Human
  • 免疫原

    Immunogen was a synthetic peptide, which represented a portion of human tripartite motif-containing 28 (KAP1) encoded within exon 1 (LocusLink ID 10155).

  • ポジティブ・コントロール

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab10483 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
WB 1/1000 - 1/20000. Detects a band of approximately 110 kDa (predicted molecular weight: 100 kDa).
IP Use at 1-4 µg/mg of lysate.
IHC-P 1/1000 - 1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC 1/500 - 1/2000.

Permeabilization with Triton-X 100 is recommended for formaldehyde-fixed cells.

ICC/IF Use at an assay dependent concentration. PubMed: 21343339
ChIP Use at an assay dependent concentration. PubMed: 17542650
ChIP/Chip Use at an assay dependent concentration. PubMed: 17542650

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 - ChIP Grade 画像

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling KAP1 with ab10483 at 1/1000 (1μg/ml). Detection: DAB.
  • Immunocytochemistry/Immunofluorescence analysis of formaldelyde-fixed HeLa cells labelling KAP1 with ab10483 at 1/1000 (1μg/ml). A FITC-conjugated goat anti-rabbit IgG (1/100) was used as the secondary antibody.


  • Predicted band size : 100 kDa

    Detection of Human KAP1 by Western Blot and Immunoprecipitation. Samples: A. Whole cell lysate (50 ug for E, 10 ug for T) from normal 293T cells (E) or 293T cells transiently transfected with a human KAP1 construct (T). B. Whole cell lysate (0.5 mg) from normal 293T cells. Antibodies: ab10483 used at the indicated concentration for WB (A) or 2 ug/0.5 mg lysate for IP (B). Immunoprecipitated KAP1 was detected using a KAP1 antibody from another source. Detection: Chemiluminescence with a 1 minute exposure.

    Detection of Human KAP1 by Western Blot and Immunoprecipitation. Samples: A. Whole cell lysate (50 ug for E, 10 ug for T) from normal 293T cells (E) or 293T cells transiently transfected with a human KAP1 construct (T). B. Whole cell lysate (0.5 mg) from normal 293T cells. Antibodies: ab10483 used at the indicated concentration for WB (A) or 2 ug/0.5 mg lysate for IP (B). Immunoprecipitated KAP1 was detected using a KAP1 antibody from another source

  • The ab10483 antibody was used to immunoprecipitate KAP1 from HEK293 nuclear extracts. Three different amounts of ab10483 or control IgG were tested (1, 2 or 4 ug). The eluates from these experiments as well as the supernatants from the KAP1 IPs were analyzed by Western blotting using the ab10483 antibody. As shown by Western blotting, the presence of a ~110 kDa band demonstrated that KAP1 was specifically precipitated from these extracts. The 50 and 25 kDa bands correspond to the IgG of the IP antibodies.

    See Abreview

  • IHC image of KAP1 staining in human liver carcinoma 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 ab10483, 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 - ChIP Grade (ab10483) 使用論文

This product has been referenced in:
  • Calderon MR  et al. Ligand-dependent corepressor contributes to transcriptional repression by C2H2 zinc-finger transcription factor ZBRK1 through association with KRAB-associated protein-1. Nucleic Acids Res 42:7012-27 (2014). ChIP . Read more (PubMed: 24829459) »
  • Castro-Diaz N  et al. Evolutionally dynamic L1 regulation in embryonic stem cells. Genes Dev 28:1397-409 (2014). Human . Read more (PubMed: 24939876) »

See all 19 Publications for this product

Product Wall

Application CHIPseq
Sample Mouse Cell lysate - nuclear (murine neuronal cell line (Neuro2a))
Specification murine neuronal cell line (Neuro2a)
Username

Abcam user community

Verified customer

投稿 Apr 15 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (hES and cancer cells)
Gel Running Conditions Reduced Denaturing (10% gel)
Loading amount 100000 cells
Specification hES and cancer cells
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C
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Abcam user community

Verified customer

投稿 Aug 20 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (mouse embryonic stem cell)
Gel Running Conditions Reduced Denaturing (10% gel)
Loading amount 100000 cells
Specification mouse embryonic stem cell
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C
Username

Abcam user community

Verified customer

投稿 Aug 20 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Liver, P5)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Na-citrate pH6
Permeabilization Yes - Triton 0,05%
Specification Liver, P5
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Sep 29 2014

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

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Thank you for contacting us.

While the exact immunogens used for these product are proprietary I am able to share these details about ab10483 and ab10484.

The epitope recognized by AB10483 maps to a region between residue 1 and 50 o...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Human Cell lysate - nuclear (Human embryonic kidney cells)
Total protein in input 100 µg
Specification Human embryonic kidney cells
Immuno-precipitation step Protein G
Username

Dr. Seth Frietze

Verified customer

投稿 Jul 04 2011

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