MDC1 Antibody

Western Blot: MDC1 Antibody [NB100-395] - Human lung cancer cell lines (UT) Untreated and (Cis) 10uM Cisplatin 16h. Image submitted by a verified customer review.

Immunocytochemistry/Immunofluorescence: MDC1 Antibody [NB100-395] - PARP activity affects 53BP1 recruitment to low-input power damage. Immunofluorescent localization of MDC1 and 53BP1 in HeLa cells irradiated with 15% (indicated by white arrowheads) and/or 30% (indicated by yellow arrowheads) at 30 min p.i. N = 10 with consistent results. Scale bar = 10 um. Image collected and cropped by CiteAb from the following publication (//academic.oup.com/nar/article-lookup/doi/10.1093/nar/gkv976), licensed under a CC-BY licence.

Western Blot: MDC1 Antibody [NB100-395] - Samples: Whole cell lysate (15 and 50 ug) from HeLa cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-MDC1 antibody NB100-395 used for WB at 0.1 ug/ml. Detection: Chemiluminescence with an exposure time of 3 minutes.

Immunoprecipitation: MDC1 Antibody [NB100-395] - Detection of human MDC1 by western blot of immunoprecipitates. Samples: Whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) from HeLa cells. Antibodies: Affinity purified rabbit anti-MDC1 antibody NB100-395 used for IP at 3 ug per reaction. MDC1 was also immunoprecipitated by two other rabbit anti-MDC1 antibodies. For blotting immunoprecipitated MDC1, NB100-395 was used at 0.4 ug/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.

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

MDC1 Antibody Summary

• Immunogen: A synthetic peptide, which represented a portion of human Mediator of DNA Damage Checkpoint Protein 1 encoded within exon 5 (NP_055456.1).
• Predicted Species: Monkey (100%), Chimpanzee (100%). Backed by our 100% Guarantee.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: MDC1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/Immunofluorescence 1:100-1:250
  • Immunoprecipitation 2 - 5 ug/mg lysate
  • Western Blot 1:2000 - 1:10000
• Application Notes: WB of lysates performed using standard western blot reagents and 4-8% SDS-PAGE. ICC/IF reactivity reported in (PMID: 26903517). MDC1 antibody validated for WB from a verified customer review.

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID: 26903517).

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 MDC1 Antibody

• DKFZp781A0122
• Em:AB023051.5
• KIAA0170MGC166888
• MDC1
• mediator of DNA damage checkpoint 1
• mediator of DNA damage checkpoint protein 1
• mediator of DNA-damage checkpoint 1
• NFBD1
• NFBD1homologue to Drosophila photoreceptor protein calphotin
• Nuclear factor with BRCT domains 1

Background

MDC1 works with H2AX to promote recruitment of repair proteins to the sites of DNA breaks and controls damage-induced cell-cycle arrest checkpoints. Cells that lack MDC1 are sensitive to ionizing radiation.

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 MDC1 Antibody (NB100-395)(18)

Publications using NB100-395

• Schleicher EM, Dhoonmoon A, Jackson LM et al. The TIP60-ATM axis regulates replication fork stability in BRCA-deficient cells Oncogenesis 2022-06-18 [PMID: 35717336] (WB, Human)
• Walter L, Canup B, Pujada A et al. Matrix metalloproteinase 9 (MMP9) limits reactive oxygen species (ROS) accumulation and DNA damage in colitis-associated cancer Cell Death Dis 2020-09-17 [PMID: 32943603] (WB, Mouse)
• Bick G, Zhang F, Meetei AR, Andreassen PR. Coordination of the recruitment of the FANCD2 and PALB2 Fanconi anemia proteins by an ubiquitin signaling network. Chromosoma 2016-06-08 [PMID: 27277787]
• Hodge CD, Ismail IH, Edwards RA et al. RNF8 E3 Ubiquitin Ligase Stimulates Ubc13 E2 Conjugating Activity That Is Essential for DNA Double Strand Break Signaling and BRCA1 Tumor Suppressor Recruitment. J Biol Chem 2016-04-29 [PMID: 26903517] (ICC/IF, Mouse)
• Ge C, Che L, Du C. The UBC Domain Is Required for BRUCE to Promote BRIT1/MCPH1 Function in DSB Signaling and Repair Post Formation of BRUCE-USP8-BRIT1 Complex. PLoS ONE. 2015-12-19 [PMID: 26683461] (ICC/IF, WB, Human)
  
  Details:
  MDC1 antibody was used for ICC/IF analysis of MDC1 in experiments involving U2OS-shBRUCE cells with stable expression of BRUCE or BRUCE mutants (Fig 6A). The antibody was also used for WB on DOX treated shBRUCE-U2OS cells which were irradiated before preparation of lysates (Fig 6C).
• Saquilabon Cruz GM, Kong X, Silva BA et al. Femtosecond near-infrared laser microirradiation reveals a crucial role for PARP signaling on factor assemblies at DNA damage sites Nucleic Acids Res 2016-02-18 [PMID: 26424850] (ICC/IF)
  
  Details:
  Potorous tridactylis (Long-nosed Potoroo)
• Samba-Louaka A, Pereira Jm, Nahori Ma et al. Listeria monocytogenes Dampens the DnA Damage Response. PLoS Pathog. 2014-10-01 [PMID: 25340842] (ICC/IF, Human)
• Silva Barbara Alcaraz, Stambaugh Jessica R, Yokomori Kyoko et al. DNA damage to a single chromosome end delays anaphase onset. J Biol Chem. 2014-08-15 [PMID: 24982423]
• Munch Sandra, Weidtkamp-Peters Stefanie, Klement Karolin et al. The tumor suppressor PML specifically accumulates at RPA/Rad51-containing DNA damage repair foci but is nonessential for DNA damage-induced fibroblast senescence. Mol Cell Biol. 2014-05-01 [PMID: 24615016]
• Mok MT, Henderson BR. The in vivo dynamic interplay of MDC1 and 53BP1 at DNA damage-induced nuclear foci. Int J Biochem Cell Biol 2012-09-01 [PMID: 22677490]
• Siddiqui-Jain A, Bliesath J, Macalino D et al. CK2 inhibitor CX-4945 suppresses DNA repair response triggered by DNA-targeted anticancer drugs and augments efficacy: mechanistic rationale for drug combination therapy. Mol Cancer Ther 2012-04-01 [PMID: 22267551]
• Reichert S, Rodel C, Mirsch J, Harter PN, Tomicic MT, Mittelbronn M, Kaina B, Rodel F. Survivin inhibition and DNA double-strand break repair: a molecular mechanism to overcome radioresistance in glioblastoma. Radiother Oncol;101(1):51-8. 2011-10-01 [PMID: 21852011] (ICC/IF, WB, Human)
• Krawczyk PM, Eppink B, Essers J et al. Mild hyperthermia inhibits homologous recombination, induces BRCA2 degradation, and sensitizes cancer cells to poly (ADP-ribose) polymerase-1 inhibition. Proc Natl Acad Sci U S A 2011-06-01 [PMID: 21555554]
• Moon SH, Lin L, Zhang X et al. Wild-type p53-induced phosphatase 1 dephosphorylates histone variant gamma-H2AX and suppresses DNA double strand break repair. J Biol Chem 2010-04-01 [PMID: 20118229]
• Shi W, Ma Z, Willers H et al. Disassembly of MDC1 Foci Is Controlled by Ubiquitin-Proteasome-dependent Degradation. J Biol Chem;283(46):31608-31616. 2008-01-01 [PMID: 18757370]
• Greubel C, Hable V, Drexler GA et al. Competition effect in DNA damage response. Radiat Environ Biophys 2008-11-01 [PMID: 18648839] (ICC/IF, Human)
• Bhoumik A, Takahashi S, Breitweiser W et al. ATM-dependent phosphorylation of ATF2 is required for the DNA damage response. Mol Cell 2005-05-01 [PMID: 15916964]
• Au WW, Henderson BR. The BRCA1 RING and BRCT domains cooperate in targeting BRCA1 to ionizing radiation-induced nuclear foci. J Biol Chem 2005-02-01 [PMID: 15569676]

Reviews for MDC1 Antibody (NB100-395) (2)

Average Rating: 4.5

(Based on 2 reviews)

We have 2 reviews tested in 2 species: Human, Other.

reviewed by: Pauline Jeannot WB Human 02/18/2019

Summary

• Application: Western Blot
• Sample Tested: human lung cancer cell line
• Species: Human
• Lot: A3

reviewed by: Barbara Alcaraz Silva IF Other 10/19/2015

Summary

• Application: Immunofluorescence
• Sample Tested: Marsupial
• Species: Other

Bioinformatics

• Gene Symbol: MDC1
• Entrez:
  • 9656(Human)
• Uniprot:
  • Q14676()