KAP1 [p Ser824] Antibody

Western Blot: KAP1 [p Ser824] Antibody [NB100-2350] - Detection of mouse Phospho KAP-1 (S824) by western blot. Samples: Whole cell lysate (50 ug) from TCMK-1 cells treated with 100 uM etoposide (+) or mock treated (-). Antibodies: Affinity purified rabbit anti-Phospho KAP-1 (S824) antibody NB100-2350 used for WB at 1 ug/ml. Detection: Chemiluminescence with an exposure time of 3 minutes.

Immunocytochemistry/Immunofluorescence: KAP1 [p Ser824] Antibody [NB100-2350] - NBF-fixed asynchronous HeLa cells grown in chambered microscope slides and treated with etoposide (left) or untreated (right). Antibody: Affinity purified rabbit anti-Phospho KAP-1 (S824) used at a dilution of 1:200 (1ug/ml). Detection: Red fluorescent Anti-rabbit IgG-DyLight (R) 594 conjugated used at a dilution of 1:100.

Immunohistochemistry-Paraffin: KAP1 [p Ser824] Antibody [NB100-2350] - Human Phospho Kap1 (S824) by Immunohistochemistry. Sample: FFPE serial section of human lung cancer used at a dilution of 1:200 (1 ug/ml). Detection: DAB

Western Blot: KAP1 [p Ser824] Antibody [NB100-2350] - Detection Human Phospho KAP-1 (S824) on HeLa whole cell lysate. KAP-1 was immunoprecipitated using another rabbit anti-KAP-1 antibody BL553.

Western Blot: KAP1 [p Ser824] Antibody [NB100-2350] - Whole cell lysate (15 and 50 ug) from 293T cells treated with 100 uM etoposide (+) or mock treated (-). Antibodies: Affinity purified rabbit anti-Phospho KAP-1 (S824) antibody used for WB at 0.1 ug/ml. Detection: Chemiluminescence with an exposure time of 30 seconds. Lower panel shows Western Blot for total KAP1-1 using affinity purified rabbit antiKAP-1 antibody at 0.1 ug/ml with an exposure time of 30 seconds.

Immunohistochemistry-Paraffin: KAP1 [p Ser824] Antibody [NB100-2350] - FFPE serial sections of asynchronous HeLa cells treated with etoposide (left) anduntreated HeLa cells (right). Antibody: Affinity purified rabbit anti-Phospho KAP-1 (S824) used at a dilution of 1:200 (1ug/ml). Detection: DAB

Immunoprecipitation: KAP1 [p Ser824] Antibody [NB100-2350] - Detection of human Phospho KAP-1 (S824) by western blot of immunoprecipitates. Samples: Whole cell lysate (1 mg for IP; 20% of IP loaded) from HEK293T cells treated with 100 uM etoposide (+) or mock treated (-). Antibodies: Rabbit anti-Phospho KAP-1 (S824) NB100-2350 and rabbit anti-KAP-1 recombinant monoclonal antibody NBP2-76393 used for IP at 6 ug/mg lysate. For blotting immunoprecipitated Phospho KAP-1 (S824), NB100-2350 was used at 0.04 ug/ml. To examine total KAP-1, the blot was stripped and then blotted with NBP2-76393 at 1:1000 (lower panel). Detection: Chemiluminescence with an exposure time of 1 second.

• Reactivity: Hu, Mu, Rt
• Applications: WB, ICC/IF, IHC, IHC-P, IP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 0.2 mg/ml

KAP1 [p Ser824] Antibody Summary

• Immunogen: Immunogen was a synthetic phosphorylated peptide, which represented a portion of human KRAB-Associated Protein 1 surrounding Serine 824 according to the numbering given in entry NP_005753.1 (GeneID 10155).
• Modification: p Ser824
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: TRIM28
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot
  • Immunocytochemistry/Immunofluorescence 1:200 - 1:1000
  • Immunohistochemistry 1:200 - 1:1000
  • Immunohistochemistry-Paraffin 1:200 - 1:1000
  • Immunoprecipitation 2-10 ug/mg lysate
• Application Notes: Epitope retrieval with citrate buffer pH 6.0 is recommended for FFPE tissue sections.
• Control:

  KAP1 Knockout HeLa Cell Lysate

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID: 24248351). Rat reactivity reported in scientific literature (PMID: 27895165).

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: TBS and 0.1% BSA
• Preservative: 0.09% Sodium Azide
• Concentration: 0.2 mg/ml
• Purity: Immunogen affinity purified

Notes

This anitbody can be used as the primary antibody in a PLA assay with the following as complementing antibodies:NB100-322, NB500-158, NB100-41429

Alternate Names for KAP1 [p Ser824] Antibody

• E3 SUMO-protein ligase TRIM28
• EC 6.3.2.-
• FLJ29029
• KAP1
• KAP-1
• KAP1KRAB-associated protein 1
• KRIP-1
• Nuclear corepressor KAP-1
• RNF96
• RNF96KRAB-interacting protein 1
• TF1B
• TIF1B
• TIF1-beta
• TIF1BRING finger protein 96
• transcription intermediary factor 1-beta
• transcriptional intermediary factor 1-beta
• TRIM28
• tripartite motif containing 28
• tripartite motif-containing 28
• Tripartite motif-containing protein 28

Background

The protein encoded by this gene mediates transcriptional control by interaction with the Kruppel-associated box repression domain found in many transcription factors. The protein localizes to the nucleus and is thought to associate with specific chromatin regions. The protein is a member of the tripartite motif family. This tripartite motif includes three zinc-binding domains, a RING, a B-box type 1 and a B-box type 2, and a coiled-coil region.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for KAP1 Antibody (NB100-2350)(30)

Publications using NB100-2350

• Quek H, Luff J, Cheung K et al. Rats with a missense mutation in Atm display neuroinflammation and neurodegeneration subsequent to accumulation of cytosolic DNA following unrepaired DNA damage. J. Leukoc. Biol. Nov 28 2016 [PMID: 27895165] (Rat)
• Hartlerode AJ, Morgan MJ, Wu Y et al. Recruitment and activation of the ATM kinase in the absence of DNA-damage sensors. Nat. Struct. Mol. Biol. 2015 Aug 17 [PMID: 26280532] (WB, Mouse)
• Kijas AW, Lim YC, Bolderson E et al. ATM-dependent phosphorylation of MRE11 controls extent of resection during homology directed repair by signalling through Exonuclease 1. Nucleic Acids Res. 2015 Aug 03 [PMID: 26240375] (WB, Human)
• Puccini J, Shalini S, Voss AK et al. Loss of caspase-2 augments lymphomagenesis and enhances genomic instability in Atm-deficient mice. Proc Natl Acad Sci U S A 2013 Dec 3 [PMID: 24248351] (WB, Mouse)
• Bhatia N, Xiao TZ, Rosenthal KA et al. MAGE-C2 Promotes Growth and Tumorigenicity of Melanoma Cells, Phosphorylation of KAP1, and DNA Damage Repair J Invest Dermatol 2012 Oct 25 [PMID: 23096706] (WB, Human)
• Yamamoto K, Wang Y, Jiang W et al. Kinase-dead ATM protein causes genomic instability and early embryonic lethality in mice. J Cell Biol 2012 Aug [PMID: 22869596]
• Bolderson E, Savage KI, Mahen R et al. Kruppel-associated Box (KRAB)-associated co-repressor (KAP-1) Ser-473 phosphorylation regulates heterochromatin protein 1beta (HP1-beta) mobilization and DNA repair in heterochromatin. J Biol Chem 2012 Aug [PMID: 22715096]
• Pires IM, Olcina MM, Anbalagan S et al. Targeting radiation-resistant hypoxic tumour cells through ATR inhibition. Br J Cancer 2012 Jul [PMID: 22713662]
• Temviriyanukul P, van Hees-Stuivenberg S, Delbos F et al. Temporally distinct translesion synthesis pathways for ultraviolet light-induced photoproducts in the mammalian genome. DNA Repair (Amst) 2012 Jun [PMID: 22521143]
• Lee DH, Goodarzi AA, Adelmant GO et al. Phosphoproteomic analysis reveals that PP4 dephosphorylates KAP-1 impacting the DNA damage response. EMBO J 2012 Apr [PMID: 22491012]
• Xiao TZ, Bhatia N, Urrutia R et al. MAGE I Transcription Factors Regulate KAP1 and KRAB Domain Zinc Finger Transcription Factor Mediated Gene Repression. PLoS ONE. 20116(8):e23747. [PMID: 21876767]
• Gatei M, Jakob B, Chen P et al. ATM protein-dependent phosphorylation of Rad50 protein regulates DNA repair and cell cycle control J Biol Chem 2011 Sep 9 [PMID: 21757780] (WB, Human)
• Sims JK, Wade PA. Mi-2/NuRD complex function is required for normal S phase progression and assembly of pericentric heterochromatin. Mol Biol Cell. 2011 Sep [PMID: 21737684]
• Nayler S, Gatei M, Kozlov S et al. Embryonic Stem Cells/Induced Pluripotent Stem (iPS) Cells: Induced Pluripotent Stem Cells from Ataxia-Telangiectasia Recapitulate the Cellular Phenotype Stem Cells Trans Med 2012 June (WB, Human)
• Du L, Kayali R, Bertoni C et al. Arginine-rich cell-penetrating peptide dramatically enhances AMO-mediated ATM aberrant splicing correction and enables delivery to brain and cerebellum Hum Mol Genet 2011 Aug 15 [PMID: 21576124] (WB, Human)
• Bhatia N, Yang B, Xiao TZ et al. Identification of novel small molecules that inhibit protein-protein interactions between MAGE and KAP-1 Arch Biochem Biophys 2011 Apr 15 [PMID: 21277283] (WB, Human)
• Forrester et al. J. Virol 85 (5):2201-2211. 2011 [PMID: 21159879]
• Jacobs et a. Hum. Mol Genet 20 (4):806-819. 2011 [PMID: 21147755]
• Kanu et al. J Biol Chem 285 (49):38534-38542. 2010 [PMID: 20889973]
• Gilmore-Hebert et al. Mol Cancer Res 8 (10):1388-1398. 2010 [PMID: 20858735]
• So et al. J Cell Biol 187 (7):977-990. 2009 [PMID: 20026654]
• Lovejoy et al. PNAS 106 (46):19304-19309. 2009 [PMID: 19889979]
• Bansbach et al. Genes & Development 23 (20):2405-2414. 2009 [PMID: 19793861]
• Chang et al. Cancer Res 69 (14):5681-5689. 2009 [PMID: 19584288]
• Tomimatsu et al. EMBO Rep 10 (6):629-635. 2009 [PMID: 19444312]
• Helmink BA, Bredemeyer AL, Lee BS et al. MRN complex function in the repair of chromosomal Rag-mediated DNA double-strand breaks. J Exp Med 2009 Mar [PMID: 19221393]
• Daniel JA, Pellegrini M, Lee JH et al. Multiple autophosphorylation sites are dispensable for murine ATM activation in vivo. J Cell Biol 2008 Dec [PMID: 19047460]
• Bencokova et al. Mol Cell Biol 29 (2):526-537. 2009 [PMID: 18981219]
• Li X, Lee YK, Jeng JC et al. Role for KAP1 serine 824 phosphorylation and sumoylation/desumoylation switch in regulating KAP1-mediated transcriptional repression. J Biol Chem 2007 Dec [PMID: 17942393]
• Biton S, Gropp M, Itsykson P et al. ATM-mediated response to DNA double strand breaks in human neurons derived from stem cells. DNA Repair (Amst) 2007 Jan [PMID: 17178256]

Bioinformatics

• Gene Symbol: TRIM28
• Entrez:
  • 10155(Human)
• Uniprot:
  • Q13263(Human)