Anti-KAP-1/TRIM28 Antibody

Transcription intermediary factor 1-beta (UniProt Q13263; also known as E3 SUMO-protein ligase TRIM28, KAP-1, KRAB-associated protein 1, KRAB-interacting protein 1, KRIP-1, Nuclear corepressor KAP-1, Protein phosphatase 1 regulatory subunit 157, RING finger protein 96, TIF1-beta, Transcriptional intermediary factor 1-beta, Tripartite motif-containing 28, Tripartite motif-containing protein 28) is encoded by the TRIM28 (also known as KAP1, PPP1R157, RNF96, TF1B, TIF1B) gene (Gene ID 10155) in human.
Kruppel-associated box-/KRAB-associated protein 1 (KAP-1) is a transcriptional co-repressor that selectively targets the KRAB domain of zinc finger protein-based transcription factors with its N-terminal tripartite RBCC (RING, b-box, coiled-coil) domain. KAP-1 uses its C-terminal PxVxL motif and the PHD-Bromo domain to recruit chromatin modifiers, such as heterochromatin protein 1 (HP-1), Histone-lysine N-methyltransferase SETDB1, nucleosome remodeling and deacetylation (NuRD) complex, and histone deacetylases (HDACs), for transcriptional repression. KAP-1 also plays an essential role in the DNA damage response (DDR). Upon DNA damage, KAP-1 is rapidly phosphorylated by Ataxia telangiectasia mutated (ATM) on Ser824, causing chromatin relaxation to facilitate DNA repair. In addition, SIRT1-mediated KAP-1 deacetylation is reported to stabilize KAP1 and 53BP1 interaction, resulting in an enhancement of NHEJ-mediated repair efficiency in a KAP-1 pSer824-independent manner.

~110 kDa observed

Epitope: Bromo domain

GST-tagged recombinant protein corresponding to the Bromo domain of human KAP-1/TRIM28.

Anti-KAP-1/TRIM28 Antibody is an antibody against KAP-1/TRIM28 for use in Western Blotting, Immunoprecipitation, Immunocytochemistry.

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Nuclear Receptors

Western Blotting Analysis: A representative lot detected the endogenous KAP-1/Trim28 only in the immunoprecipitats of exogenously expressed FLAG-SirT1 constructs that contain the KAP-1/Trim28-binding domain (a.a. 143-213) using various transfected HEK293T cells (Lin, Y.H., et al. (2015). PLoS One. 10(4):e0123935).
Western Blotting Analysis: A representative lot detected similar level of endogenous KAP-1/Trim28 in HEK293T cells regardless of ionizing radiation treatment or shRNA-mediated SirT1 downregulation (Lin, Y.H., et al. (2015). PLoS One. 10(4):e0123935).
Western Blotting Analysis: A representative lot detected KAP-1/Trim28 in whole cell lysates, but not in BAP1, FoxK1, or FoxK2 immunoprecipitate (Okino, Y., et al. (2015). J. Biol. Chem. 16;290(3):1580-1591).
Immunoprecipitation Analysis: 10 µL from a representative lot immunoprecipitated KAP-1/Trim28 from HEK293T cell lysates (Courtesy of Dr. Zhenkun Lou, Mayo clinic, Rochester, MN).
Immunocytochemistry Analysis: An 1:1000 dilution from a representative lot detected KAP-1/Trim28 nuclear localization in U2OS cells (Courtesy of Dr. Zhenkun Lou, Mayo clinic, Rochester, MN).
Immunoprecipitation Analysis: A representative lot co-immunoprecipitated SirT1 with KAP-1/Trim28 from HEK293T cell lysates (Lin, Y.H., et al. (2015). PLoS One. 10(4):e0123935).
Western Blotting Analysis: An 1:1000 dilution from a representative lot detected KAP-1/Trim28 in mouse embryonic fibroblast (MEF) lysate, as well as downregulated KAP-1/Trim28 in lysates from KAP1 siRNA-treated HEK293T cells (Courtesy of Dr. Zhenkun Lou, Mayo clinic, Rochester, MN).

Expected to react with both spliced isoforms.

Rabbit polyclonal antibody in serum with 0.05% sodium azide.

Unpurified

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Evaluated by Western Blotting in HEK293T cell lysate.

Western Blotting Analysis: A 1:2,000 dilution of this antibody detected KAP-1/TRIM28 in 10 µg of HEK293T cell lysate.

Concentration: Please refer to lot specific datasheet.

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.