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Overview
Product name
Anti-KAP1 antibody - ChIP Grade
See all KAP1 products (16) ...
Description
Rabbit polyclonal to KAP1 - ChIP Grade
Tested applications
WB, IP, ICC/IF, ChIP, ChIP/Chipmore details
Cross reactivity
Reacts with
Mouse, Human
Immunogen
Immunogen was a synthetic peptide, which represented a portion of human tripartite motif-containing 28 (KAP1) encoded within exon 1 (LocusLink ID 10155).
Positive control
Whole cell lysate from normal 293T cells or 293T cells transiently transfected with a human KAP1 construct.
Properties
Form
Liquid
Storage instructions
Store at +4°C.
Storage buffer
Preservative: 0.1% Sodium Azide
Constituents: 8mM PBS, 60mM Citrate, 150mM Tris, pH 7-8
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Purification notes
Antibodies were affinity purified using the peptide immobilized on solid support.
Clonality
Polyclonal
Isotype
IgG
Research areas
Epigenetics and Nuclear Signaling >> ChIP'ing antibodies >> ChIP'ing antibodies
Epigenetics and Nuclear Signaling >> Transcription >> Transcription Factors
Epigenetics and Nuclear Signaling >> Transcription >> Other factors
Cell Biology >> Proteolysis / Ubiquitin >> Proteasome / Ubiquitin >> Ubiquitin E2s and E3s >> RING Finger E3 Ligase
Epigenetics and Nuclear Signaling >> Chromatin Binding Proteins >> Methylated DNA
Applications
Show applications key
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.
WB: 1/500 - 1/2000.Detects a band of approximately 110 kDa (predicted molecular weight: 100 kDa).
IP: Use a concentration of 1 - 4 µg/ml.
ICC/IF: Use at an assay dependent dilution. (PubMed: 21343339)
ChIP: Use at an assay dependent dilution. (PubMed: 17542650)
ChIP/Chip: Use at an assay dependent dilution. (PubMed: 17542650)
Target
Function
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.
Tissue specificity
Expressed in all tissues tested including spleen, thymus, prostate, testis, ovary, small intestine, colon and peripheral blood leukocytes.
Pathway
Protein modification; protein sumoylation.
Sequence similarities
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.
Domain
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.
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 localization
Nucleus. Associated with centromeric heterochromatin during cell differentiation through CBX1.
Target information above from: UniProt accessionQ13263
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- E3 SUMO-protein ligase TRIM28 antibody
- FLJ29029 antibody
- KAP 1 antibody
see all
Database links
- 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
Anti-KAP1 antibody - ChIP Grade images:
Western blot - KAP1 antibody (ab10483)
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.
Immunoprecipitation - Anti-KAP1 antibody - ChIP Grade (ab10483)
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.
This image was taken from an abreview submitted by an Seth Frietze.
References for Anti-KAP1 antibody - ChIP Grade (ab10483)
This product has been referenced in:
- Iyengar Set al. Functional analysis of KAP1 genomic recruitment. Mol Cell Biol 31:1833-47 (2011). ICC/IF, ChIP; Human.Read more (PubMed: 21343339) »
- Frietze Set al. ZNF274 recruits the histone methyltransferase SETDB1 to the 3' ends of ZNF genes. PLoS One 5:e15082 (2010). ChIP/Chip; Human.Read more (PubMed: 21170338) »
See all 5 publications for this product
Publishing research using ab10483? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"