Anti-KAP1 (phospho S824) antibody (ab70369)

Overview

  • Product nameAnti-KAP1 (phospho S824) antibody
    See all KAP1 primary antibodies
  • Description
    Rabbit polyclonal to KAP1 (phospho S824)
  • Tested applicationsSuitable for: ICC, WB, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Chimpanzee, Rhesus monkey, Gorilla, Orangutan
  • Immunogen

    A synthetic phosphorylated peptide, corresponding to a sequence of human KAP1 surrounding Serine 824. NP_005753.1

  • Positive control
    • Whole cell lysate of asynchronous HeLa cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab70369 in the following tested applications.

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

Application Abreviews Notes
ICC Use at an assay dependent concentration.
WB 1/1000 - 1/5000. Detects a band of approximately 117 kDa (predicted molecular weight: 89 kDa).
ICC/IF 1/1000.
IP Use at 2-5 µg/mg of lysate.

Target

  • FunctionNuclear 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.
  • Tissue specificityExpressed in all tissues tested including spleen, thymus, prostate, testis, ovary, small intestine, colon and peripheral blood leukocytes.
  • PathwayProtein modification; protein sumoylation.
  • Sequence similaritiesBelongs 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.
  • DomainThe 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.
  • Post-translational
    modifications
    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.
  • Cellular localizationNucleus. Associated with centromeric heterochromatin during cell differentiation through CBX1.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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 (phospho S824) antibody images

  • All lanes : Anti-KAP1 (phospho S824) antibody (ab70369) at 0.1 µg/ml

    Lane 1 : Whole cell lysate from asynchronous HeLa cells, treated with neocarzinostatin (NCS; 200 ng/ml, 30 minutes) at 50 µg
    Lane 2 : Whole cell lysate from asynchronous HeLa cells at 50 µg
    Lane 3 : Whole cell lysate from asynchronous HeLa cells, treated with neocarzinostatin (NCS; 200 ng/ml, 30 minutes) at 200 µg
    Lane 4 : Whole cell lysate from asynchronous HeLa cells at 200 µg

    Developed using the ECL technique

    Predicted band size : 89 kDa
    Observed band size : 117 kDa (why is the actual band size different from the predicted?)
  • ab70369 staining KAP1 (phospho S824) (red) in Human U2OS cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were treated with 1 µM Etoposide for 3 hours prior to fixing. Cells were fixed with formaldehyde, permeabilized in 0.5% NP40 and blocked with 3% BSA for 2 hours at 21°C. Samples were incubated with primary antibody (1/1000 in PBS + 3% BSA) for 12 hours at 4°C. An Alexa Fluor®594-conjugated Mouse anti-rabbit IgG monoclonal (1/500) was used as the secondary antibody.

    See Abreview

References for Anti-KAP1 (phospho S824) antibody (ab70369)

This product has been referenced in:

See all 8 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (vascular smooth muscle cell)
Permeabilization Yes - NP40
Specification vascular smooth muscle cell
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3µg/mL · Temperature: 22°C
Fixative Formaldehyde
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Submitted Nov 03 2015

Application Immunocytochemistry
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
Sample Human Cultured Cells (U2OS)
Specification U2OS
Permeabilization Yes - 0.5% NP40
Fixative Paraformaldehyde
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Submitted Oct 02 2013

Merci de votre intérêt pour ab70369.

Effectivement, cet anticorps n’a pas encore été testé chez la souris et nous n’offrons pas d’échantillon gratuit à des fins de tests pr&eac...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (u2os)
Specification u2os
Fixative Formaldehyde
Permeabilization Yes - 0.5% NP40
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
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Ms. Can Zhou

Verified customer

Submitted May 21 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - nuclear (U20S)
Loading amount 12 µg
Specification U20S
Treatment 1 uM Doxorubicin for stated times (in mins)
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
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Submitted May 05 2012

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