Anti-KAP1 antibody (ab10483)
Key features and details
- Rabbit polyclonal to KAP1
- Suitable for: WB, IHC-P, IP
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Overview
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Product name
Anti-KAP1 antibody
See all KAP1 primary antibodies -
Description
Rabbit polyclonal to KAP1 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. The immunogen is between aa 1-50.
Database link: Q13263 -
Positive control
- WB: human HeLa, HEK293T cells; mouse NIH3T3, TCMK-1, 4T1, CT26.WT, rat C6 cells. IP: HeLa cells. IHC: human breast carcinoma
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
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Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7
Preservative: 0.1% Sodium azide
Constituents: 0.021% PBS, 1.764% Sodium citrate, 1.815% Tris -
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
Associated products
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ChIP Related Products
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
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.
| Application | Abreviews | Notes |
|---|---|---|
| WB | (3) |
1/2000 - 1/10000. Detects a band of approximately 110 kDa (predicted molecular weight: 100 kDa).
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| IHC-P | (1) |
1/500 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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| IP | (1) |
Use at 2-10 µg/mg of lysate.
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| Notes |
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WB
1/2000 - 1/10000. Detects a band of approximately 110 kDa (predicted molecular weight: 100 kDa). |
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IHC-P
1/500 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
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IP
Use at 2-10 µg/mg of lysate. |
Target
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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
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 localization
Nucleus. Associated with centromeric heterochromatin during cell differentiation through CBX1. - Information by UniProt
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Database links
- Entrez Gene: 10155 Human
- Entrez Gene: 21849 Mouse
- Entrez Gene: 116698 Rat
- Omim: 601742 Human
- SwissProt: Q13263 Human
- SwissProt: Q62318 Mouse
- SwissProt: O08629 Rat
- Unigene: 467408 Human
see all -
Alternative names
- E3 SUMO protein ligase TRIM28 antibody
- E3 SUMO-protein ligase TRIM28 antibody
- FLJ29029 antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAP1 antibody (ab10483)
Immunohistochemistry analysis of formalin-fixed paraffin embedded sections of human breast tissue staining KAP-1 with ab10483 at 1/1000 dilution, showing the current lot on the left, and previous lot on the right.
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Immunoprecipitation - Anti-KAP1 antibody (ab10483)Immunoprecipitation of KAP-1 from HeLa whole cell lysate using ab10483
Lane 1: ab10483 current lot
Lane 2: ab10483 previous lot
Lane 3: other rabbit anti-KAP-1 antibody
Lane 4: Control IgG
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Western blot - Anti-KAP1 antibody (ab10483)All lanes : Anti-KAP1 antibody (ab10483) at 1 µg/ml
Lane 1 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate
Lane 2 : TCMK-1 (mouse kidney epithelial cell line) whole cell lysate
Lane 3 : 4T1 (Mouse mammary gland carcinoma cell line) whole cell lysate
Lane 4 : CT26.WT (murine colon carcinoma) whole cell lysate
Lane 5 : C6 (rat glioma cell line) whole cell lysate
Lysates/proteins at 50 µg/ml per lane.
Predicted band size: 100 kDa
Exposure time: 3 minutes -
Western blot - Anti-KAP1 antibody (ab10483)All lanes : Anti-KAP1 antibody (ab10483) at 0.1 µg/ml
Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 50 µg
Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 15 µg
Lane 3 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 5 µg
Lane 4 : HEK293T cells (human epithelial cell line from embryonic kidney transformed with large T antigen) at 50 µg
Predicted band size: 100 kDa
Exposure time: 30 seconds -
Immunoprecipitation - Anti-KAP1 antibody (ab10483)This image was taken from an abreview submitted by an Seth Frietze.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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (66)
ab10483 has been referenced in 66 publications.
- Collier AJ et al. Genome-wide screening identifies Polycomb repressive complex 1.3 as an essential regulator of human naïve pluripotent cell reprogramming. Sci Adv 8:eabk0013 (2022). PubMed: 35333572
- Acurzio B et al. The mismatch-repair proteins MSH2 and MSH6 interact with the imprinting control regions through the ZFP57-KAP1 complex. Epigenetics Chromatin 15:27 (2022). PubMed: 35918739
- Fraser CR et al. Radiofluorination of a highly potent ATM inhibitor as a potential PET imaging agent. EJNMMI Res 12:50 (2022). PubMed: 35962885
- Ait-Ammar A et al. Inhibition of HIV-1 gene transcription by KAP1 in myeloid lineage. Sci Rep 11:2692 (2021). PubMed: 33514850
- Diao Z et al. SIRT3 consolidates heterochromatin and counteracts senescence. Nucleic Acids Res 49:4203-4219 (2021). PubMed: 33706382