Product nameAnti-KAP1 antibody [20C1] - ChIP Grade
See all KAP1 primary antibodies
DescriptionMouse monoclonal [20C1] to KAP1 - ChIP Grade
Tested applicationsSuitable for: ChIP, IHC-P, ICC/IF, WB, EMSA, IPmore details
Species reactivityReacts with: Mouse, Rat, Human
Recombinant fragment, corresponding to amino acids 60-383 of Human KAP1.
- Purchase matching WB positive control:Recombinant Human KAP1 protein
- Gamma irradiated HeLa cell lysate (WB), and cells (ICC). This antibody gave a positive result in IHC in the following FFPE tissue: Human spleen.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.05% Sodium azide
Constituents: PBS, 0.1% BSA
Concentration information loading...
PurityProtein G purified
- Cell Biology
- Proteolysis / Ubiquitin
- Proteasome / Ubiquitin
- Ubiquitin E3 Enzymes
- RING Finger E3 Ligase
ChIP Related Products
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.
|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.|
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.
modificationsPhosphorylated 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
- E3 SUMO protein ligase TRIM28 antibody
- E3 SUMO-protein ligase TRIM28 antibody
- FLJ29029 antibody
Anti-KAP1 antibody [20C1] - ChIP Grade (ab22553) at 1/500 dilution + HeLa whole cell lysate
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
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.
This product has been referenced in:
- Lee JH et al. ATM directs DNA damage responses and proteostasis via genetically separable pathways. Sci Signal 11:N/A (2018). Read more (PubMed: 29317520) »
- Kato M et al. A somatic role for the histone methyltransferase Setdb1 in endogenous retrovirus silencing. Nat Commun 9:1683 (2018). Read more (PubMed: 29703894) »