TopBP1 Antibody

Western Blot: TopBP1 Antibody [NB100-217] - Detection of mouse TopBP1 by western blot. Samples: Whole cell lysate (50 ug) from TCMK-1 and NIH 3T3 cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-TopBP1 antibody NB100-217 used for WB at 0.1 ug/ml. Detection: Chemiluminescence with an exposure time of 3 minutes.

Western Blot: TopBP1 Antibody [NB100-217] - Sample: Whole cell lysate (50 ug for WB) from HeLa cells.

Western Blot: TopBP1 Antibody [NB100-217] - WB analysis of TOPBP1 in HeLa cell extract mitotic and interphase. Image courtesy of product review submitted by Jennifer Flynn.

Western Blot: TopBP1 Antibody [NB100-217] - Detection of Mouse. Whole cell lysate from TCMK-1 (15 and 50 ug) cells.

Western Blot: TopBP1 Antibody [NB100-217] - Samples: Whole cell lysate (50 ug) from HeLa and 293T cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-TOPBP1 antibody used for WB at 0.1 ug/ml. Detection: Chemiluminescence with an exposure time of 3 minutes.

Western Blot: TopBP1 Antibody [NB100-217] - Detection of human TopBP1 by western blot. Samples: Whole cell lysate (15 ug) from HeLa, HEK293T, and Jurkat cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-TopBP1 antibody NB100-217 used for WB at 0.1 ug/ml. Detection: Chemiluminescence with an exposure time of 10 seconds.

Immunoprecipitation: TopBP1 Antibody [NB100-217] - Detection of human TopBP1 by western blot of immunoprecipitates. Samples: Whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) from HeLa cells prepared using NETN lysis buffer. Antibodies: Affinity purified rabbit anti-TopBP1 antibody NB100-217 (lot NB100-217-2) used for IP at 3 ug per reaction. TopBP1 was also immunoprecipitated by a previous lot of this antibody (lot A300-111A-1). For blotting immunoprecipitated TopBP1, A300-111A was used at 0.4 ug/ml. Detection: Chemiluminescence with an exposure time of 10 seconds.

• Reactivity: Hu, Mu
• Applications: WB, IP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

TopBP1 Antibody Summary

• Immunogen: The immunogen recognized by this antibody maps to a region between residue 1400 and the C-terminus (residue 1435) of human Topoisomerase (DNA) II Binding Protein 1 using the numbering given in entry NP_008958.1 (GeneID 11073).
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: TOPBP1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot
  • Immunoprecipitation 0.5 to 2.5 ug/mg of lysate

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: Tris-Citrate/Phosphate (pH 7.0 - 8.0)
• Preservative: 0.09% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for TopBP1 Antibody

• DNA topoisomerase II-beta-binding protein 1
• DNA topoisomerase II-binding protein 1
• KIAA0259DNA topoisomerase 2-binding protein 1
• TOP2BP1
• TopBP1
• topoisomerase (DNA) II binding protein 1

Background

Topoisomerases regulate topological states of DNA. DNA topoisomerase II-binding protein 1 (TopBP1) is a protein that interacts with the C-terminal region of topoisomerase II beta (Topo II beta). TopBP1 function has been found to be important to various aspects of cellular growth control such as DNA replication, the DNA damage checkpoint, and cell survival. TopBP1 interacts with several proteins through its BRCA1 C-terminal (BRCT) motifs. TopBP1 interacts and inhibits E2F1 transcriptional activity through an Rb-independent mechanism that involves recruitment of the SWI/SNF chromatin-remodeling complex proteins Brg1 and Brm. TopBP1 is also known as DNA topoisomerase 2-binding protein, DNA topoisomerase II beta-binding protein 1, TOPBP1, and KIAA0259.

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 TopBP1 Antibody (NB100-217)(18)

Publications using NB100-217

• Choe KN, Nicolae CM, Constantin D et al. HUWE1 interacts with PCNA to alleviate replication stress. EMBO Rep. May 4 2016 [PMID: 27146073] (WB, Human)
• Tomida J, Itaya A, Shigechi T et al. A novel interplay between the Fanconi anemia core complex and ATR-ATRIP kinase during DNA cross-link repair. Nucleic Acids Res 2013 May 30 [PMID: 23723247] (WB, Human)
• Ohlenschlager O, Kuhnert A, Schneider A et al. The N-terminus of the human RecQL4 helicase is a homeodomain-like DNA interaction motif Nucleic Acids Res 2012 Jun 22 [PMID: 22730300] (IP, WB, Human)
• Lee Y, Katyal S, Downing SM et al. Neurogenesis requires TopBP1 to prevent catastrophic replicative DNA damage in early progenitors. Nat Neurosci 2012 Jun [PMID: 22522401]
• Kuhnert A, Schmidt U, Monajembashi S et al. Proteomic identification of PSF and p54(nrb) as topBP1-interacting proteins J. Cell. Biochem 2012 [PMID: 22213094] (WB, Human)
• Kumagai A, Shevchenko A, Shevchenko A et al. Direct regulation of Treslin by cyclin-dependent kinase is essential for the onset of DNA replication. J Cell Biol 2011 Jun [PMID: 21646402]
• Forrester NA, Sedgwick GG, Thomas A et al. Serotype-specific inactivation of the cellular DNA damage response during adenovirus infection. J Virol 2011 Mar [PMID: 21159879]
• Blackford AN, Patel RN, Forrester NA et al. Adenovirus 12 E4orf6 inhibits ATR activation by promoting TOPBP1 degradation. Proc Natl Acad Sci U S A 2010 Jul [PMID: 20566845]
• Guo R, Chen J, Zhu F et al. E2F1 localizes to sites of UV-induced DNA damage to enhance nucleotide excision repair. J Biol Chem 2010 Jun [PMID: 20413589]
• Sansam CL, Cruz NM, Danielian PS et al. A vertebrate gene, ticrr, is an essential checkpoint and replication regulator. Genes Dev 2010 Jan 15 [PMID: 20080954] (WB, Human)
• Liu Y, Fang Y, Shao H et al. Interactions of human mismatch repair proteins MutSalpha and MutLalpha with proteins of the ATR-Chk1 pathway. J Biol Chem 2010 Feb [PMID: 20029092]
• Liu K, Bellam N, Lin HY et al. Regulation of p53 by TopBP1: a potential mechanism for p53 inactivation in cancer. Mol Cell Biol 2009 May [PMID: 19289498]
• Xu X, Vaithiyalingam S, Glick GG et al. The basic cleft of RPA70N binds multiple checkpoint proteins, including RAD9, to regulate ATR signaling. Mol Cell Biol 2008 Dec [PMID: 18936170]
• Hein J, Boichuk S, Wu J et al. Simian virus 40 large T antigen disrupts genome integrity and activates a DNA damage response via Bub1 binding. J Virol 2009 Jan [PMID: 18922873]
• Delacroix S, Wagner JM, Kobayashi M et al. The Rad9-Hus1-Rad1 (9-1-1) clamp activates checkpoint signaling via TopBP1. Genes Dev 2007 Jun [PMID: 17575048]
• Zhang D, Zaugg K, Mak TW et al. A role for the deubiquitinating enzyme USP28 in control of the DNA-damage response. Cell 2006 Aug [PMID: 16901786]
• Liu K, Luo Y, Lin FT et al. TopBP1 recruits Brg1/Brm to repress E2F1-induced apoptosis, a novel pRb-independent and E2F1-specific control for cell survival. Genes Dev 2004 Mar [PMID: 15075294]
• Dhoonmoon A, Schleicher EM, Nicolae CM Genome-wide CRISPR synthetic lethality screen identifies a role for the ADP-ribosyltransferase PARP14 in replication fork stability controlled by ATR bioRxiv Jan 1 2020 (WB, Human)

Review for TopBP1 Antibody (NB100-217) (1)

Average Rating: 3

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Anonymous WB Human 07/21/2010

Summary

• Application: Western Blot
• Sample Tested: HeLa Cell Extract - Mitotic and Interphase, Sample Amount: Approximately 5x10^6 cell equivalents/lane
• Species: Human
• Comments: The antibody worked to detect a protein the size of TopBP1 (~180kDa) in Hela cell extract (interphase and mitotic) but also recognized a band of a smaller size (~95kDa) and with much more intensity.

Blocking

• Blocking Details: Blocking Buffer: 5% non-fat dry milk in PBST, Blocking Time: 45 minutes, Blocking Temp: Room temperature

Primary Anitbody

• Dilution Ratio: Dilution Ratio: 1:2,000, Incubation Dilution Buffer: 5% non-fat dry milk in PBST, Incubation Time: 15 hours, Incubation Temp: 4C

Secondary Antibody

• Secondary Description: Secondary Ab: Goat-anti-Rb-HRP in 5% non-fat dry milk/PBST, Secondary Ab Dilution Ratio: 1:20,000
• Secondary Manufacturer Cat#: Jackson ImmunoResearch Cat #111-035-144

Details

• Detection Notes: Method: SuperSignal West Pico Chemiluminescent Substrate, Exposure Time: 5 min, Bands: 180 kDa(specific), 95 kDa(non-specific)

Comments

• Comments: The antibody worked to detect a protein the size of TopBP1 (~180kDa) in Hela cell extract (interphase and mitotic) but also recognized a band of a smaller size (~95kDa) and with much more intensity.

Bioinformatics

• Gene Symbol: TOPBP1
• Entrez:
  • 11073(Human)
• Uniprot:
  • Q92547(Human)