Overview

  • Product name
  • Description
    Rabbit polyclonal to MDC1
  • Tested applications
    Suitable for: ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Chimpanzee, Rhesus monkey, Gorilla
  • Immunogen

    Synthetic peptide, which represents a portion of the human Mediator of DNA Damage Checkpoint Protein 1 encoded within exon 5 (LocusLink ID 9656).

  • Positive control
    • Tested with HeLa cells and HEK 293 cells.
  • General notes

    In response to recent feedback about mouse species we no longer guarantee this anymore. Please do contact our scientific support team for more information.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.1% Sodium Azide
    Constituents: 8mM PBS, 60mM Citrate, 150mM Tris, pH 7-8
  • 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

Applications

Our Abpromise guarantee covers the use of ab11169 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
ICC/IF 1/100 - 1/250.
WB 1/500 - 1/2500. Detects a band of approximately 250 kDa (predicted molecular weight: 220 kDa).

Target

  • Function
    Required for checkpoint mediated cell cycle arrest in response to DNA damage within both the S phase and G2/M phases of the cell cycle. May serve as a scaffold for the recruitment of DNA repair and signal transduction proteins to discrete foci of DNA damage marked by 'Ser-139' phosphorylation of histone H2AFX. Also required for downstream events subsequent to the recruitment of these proteins. These include phosphorylation and activation of the ATM, CHEK1/CHK1 and CHEK2/CHK2/CDS1 kinases, and stabilization of TP53 and apoptosis. ATM and CHEK2 may also be activated independently by a parallel pathway mediated by TP53BP1.
  • Tissue specificity
    Highly expressed in testis.
  • Sequence similarities
    Contains 2 BRCT domains.
    Contains 1 FHA domain.
  • Domain
    Tandemly repeated BRCT domains are characteristic of proteins involved in DNA damage signaling. In MDC1, these repeats are required for localization to chromatin which flanks sites of DNA damage marked by 'Ser-139' phosphorylation of H2AFX.
  • Post-translational
    modifications
    Phosphorylated upon exposure to ionizing radiation (IR), ultraviolet radiation (UV), and hydroxyurea (HU). Phosphorylation in response to IR requires ATM, NBN, and possibly CHEK2. Also phosphorylated during the G2/M phase of the cell cycle and during activation of the mitotic spindle checkpoint.
  • Cellular localization
    Nucleus. Associated with chromatin. Relocalizes to discrete nuclear foci following DNA damage, this requires 'Ser-139' phosphorylation of H2AFX. Colocalizes with APTX at sites of DNA double-strand breaks.
  • Information by UniProt
  • Database links
  • Alternative names
    • Homologue to Drosophila photoreceptor protein calphotin antibody
    • MDC 1 antibody
    • Mdc1 antibody
    • MDC1_HUMAN antibody
    • Mediation of DNA damage checkpoint 1 antibody
    • Mediator of DNA damage checkpoint 1 antibody
    • Mediator of DNA damage checkpoint protein 1 antibody
    • NFBD 1 antibody
    • NFBD1 antibody
    • Nuclear factor with BRCT domains 1 antibody
    • Nuclear Factor with BRCT Domains Protein 1 antibody
    see all

Images

  • ab11169 staining MDC1 in HCT116 cells treated with GANT 61 (ab120904), by ICC/IF. Increase in MDC1 expression correlates with increased concentration of GANT 61, as described in literature.
    The cells were incubated at 37°C for 4 hours in media containing different concentrations of ab120904 (GANT 61) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab11169 (1/200 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.


  • Predicted band size : 220 kDa

    Samples: A) Nuclear extract (50 mcg) from HeLa cells. B) Whole cell lysate from HEK 293 cells. Antibodies: A) ab11169 used at the indicated concentrations for WB. B) ab11170 (column labelled BL579) and ab11171 (column labelled BL580) (a ) used at 3.3 mcg/mg lysate for IP followed by WB using ab11169 at 0.1 mcg/ml. Detection: Chemiluminescence with exposure times less than 5 min.

    Samples: A) Nuclear extract (50 mcg) from HeLa cells. B) Whole cell lysate from HEK 293 cells. Antibodies: A) ab11169 used at the indicated concentrations for WB. B) ab11170 (column labelled BL579) and ab11171 (column labelled BL580) (a ) used at 3.3 mcg/mg lysate for IP followed by WB using ab11169 at 0.1 mcg/ml. Detection: Chemiluminescence with exposure times less than 5 min.

  • ab11169 staining MDC1 in human cervical cancer cells (HeLa) by ICC/IF (immunocytochemistry/immunofluorescence - left image). Cells were paraformaldehyde fixed, permeabilized with Triton X-100 and blocked with 5% BSA for 1 hour at 25°C. The primary antibody (1/200) was incubated with the sample for 12 hours at 4°C. An Alexa Fluor 594®-conjugated Chicken anti-rabbit polyclonal (1/2000) was used as the secondary. The nuclei were stained with DAPI (right image).

    See Abreview

  • Immunofluorescence analysis of A549 cells before (left) and after ionizing radiation (right), staining MDC1 with ab11169.

    Cells were fixed with paraformaldehyde, permeabilized with 0.2% Triton X-100 and blocked with 1% BSA for 30 minutes at room temperature. Cells were incubated with primary antibody for 1 hour at room temperature. A Cy2®-conjugated anti-rabbit IgG was used as the secondary antibody.

References

This product has been referenced in:
  • Cristini A  et al. DNA-PK triggers histone ubiquitination and signaling in response to DNA double-strand breaks produced during the repair of transcription-blocking topoisomerase I lesions. Nucleic Acids Res 44:1161-78 (2016). ICC/IF ; Human . Read more (PubMed: 26578593) »
  • Renaud E  et al. Impaired TIP60-mediated H4K16 acetylation accounts for the aberrant chromatin accumulation of 53BP1 and RAP80 in Fanconi anemia pathway-deficient cells. Nucleic Acids Res 44:648-56 (2016). ICC/IF ; Human . Read more (PubMed: 26446986) »

See all 28 Publications for this product

Customer reviews and Q&As

Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293)
Gel Running Conditions
Reduced Denaturing
Loading amount
50 µg
Treatment
IR
Specification
HEK293
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Oct 26 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - 0.2% Triton X-100
Specification
HeLa
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
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Submitted Oct 06 2016

Abreviews
Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing (7.5%)
Sample
Mouse Cell lysate - other (MEF)
Specification
MEF
Treatment
with/without IR 20Gy 1hr
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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Verified customer

Submitted Feb 25 2015

Application
IHC - Wholemount
Sample
Mouse Embryo (stage e6.5)
Specification
stage e6.5
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Submitted Dec 15 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Sample
Human Cell (U2OS osteosarcoma)
Specification
U2OS osteosarcoma
Permeabilization
Yes - 0.5% Triton CSK buffer pre-extraction
Fixative
Formaldehyde
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Submitted Mar 31 2014

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (6%)
Sample
Human Cell lysate - whole cell (U2OS osteosarcoma)
Specification
U2OS osteosarcoma
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Submitted Mar 31 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (cervical cancer)
Specification
cervical cancer
Fixative
Paraformaldehyde
Permeabilization
Yes - Triton X-100
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Verified customer

Submitted Dec 02 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - nuclear (Hek293, MCF-7 and rat Vascular Smooth Muscle Cells)
Loading amount
20 µg
Specification
Hek293, MCF-7 and rat Vascular Smooth Muscle Cells
Gel Running Conditions
Reduced Denaturing (5% gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

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Verified customer

Submitted Sep 22 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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