Overview

  • Product name

    Anti-Rad50 antibody [EPR20968] - ChIP Grade
    See all Rad50 primary antibodies
  • Description

    Rabbit monoclonal [EPR20968] to Rad50 - ChIP Grade
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ChIP, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human Rad50 aa 1-450. The exact sequence is proprietary.
    Database link: Q92878

  • Positive control

    • WB: Human fetal brain and kidney lysates; K562, HeLa, HEK-293T, HT-29, NIH/3T3, ES-D3, C6, RAW 264.7 and PC-12 cell lysates; Mouse kidney lysate; Rat brain and spleen lysate. IHC-P: Human breast and breast carcinoma tissue; Mouse and rat testis tissue. ChIP: Chromatin prepared from HeLa cells.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab208019 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
WB 1/1000. Detects a band of approximately 154 kDa (predicted molecular weight: 153 kDa).
ChIP Use 2 µg for 25 µg of chromatin.
IHC-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Function

    Component of the MRN complex, which plays a central role in double-strand break (DSB) repair, DNA recombination, maintenance of telomere integrity and meiosis. The complex possesses single-strand endonuclease activity and double-strand-specific 3'-5' exonuclease activity, which are provided by MRE11A. RAD50 may be required to bind DNA ends and hold them in close proximity. This could facilitate searches for short or long regions of sequence homology in the recombining DNA templates, and may also stimulate the activity of DNA ligases and/or restrict the nuclease activity of MRE11A to prevent nucleolytic degradation past a given point. The complex may also be required for DNA damage signaling via activation of the ATM kinase. In telomeres the MRN complex may modulate t-loop formation.
  • Tissue specificity

    Expressed at very low level in most tissues, except in testis where it is expressed at higher level. Expressed in fibroblasts.
  • Involvement in disease

    Defects in RAD50 are the cause of Nijmegen breakage syndrome-like disorder (NBSLD) [MIM:613078]; also called NBS-like disorder or RAD50 deficiency. NBSLD is a disorder similar to Nijmegen breakage syndrome and characterized by chromosomal instability, radiation sensitivity, microcephaly, growth retardation, short stature and bird-like face. Immunodeficiency is absent.
  • Sequence similarities

    Belongs to the SMC family. RAD50 subfamily.
    Contains 1 zinc-hook domain.
  • Domain

    The zinc-hook, which separates the large intramolecular coiled coil regions, contains 2 Cys residues that coordinate one molecule of zinc with the help of the 2 Cys residues of the zinc-hook of another RAD50 molecule, thereby forming a V-shaped homodimer. The two heads of the homodimer, which constitute the ATP-binding domain, interact with the MRE11A homodimer.
  • Post-translational
    modifications

    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Cellular localization

    Nucleus. Chromosome > telomere. Localizes to discrete nuclear foci after treatment with genotoxic agents.
  • Information by UniProt
  • Database links

  • Alternative names

    • DNA repair protein RAD50 antibody
    • hRAD50 antibody
    • hsRAD50 antibody
    • Mrell antibody
    • NBSLD antibody
    • Rad 50 antibody
    • RAD50 2 antibody
    • rad50 antibody
    • RAD50 homolog (S cerevisiae) antibody
    • RAD50 homolog antibody
    • RAD50 PEN antibody
    • RAD50, S. cerevisiae, homolog of antibody
    • RAD50_HUMAN antibody
    • RAD502 antibody
    • Rad50l antibody
    • Truncated RAD50 protein antibody
    see all

Images

  • All lanes : Anti-Rad50 antibody [EPR20968] - ChIP Grade (ab208019) at 1/1000 dilution

    Lane 1 : Mouse kidney lysate at 10 µg
    Lane 2 : Rat brain lysate at 10 µg
    Lane 3 : Rat spleen lysate at 10 µg
    Lane 4 : C6 (rat glial tumor cell line) whole cell lysate at 20 µg
    Lane 5 : RAW 364.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 20 µg
    Lane 6 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate at 20 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 153 kDa
    Observed band size: 154 kDa
    why is the actual band size different from the predicted?



    Blocking/Diluting buffer and concentration: 5% NFDM/TBST.

    Exposure time: Lanes 1-3: 3 minutes; Lanes 4-6: 41 seconds.

    The expression profile observed is consistent with what has been described in the literature (PMID: 26068589).

  • Chromatin was prepared from HeLa (human epithelial cell line from cervix adenocarcinoma) cells treated with 1% methymethanesulfonate (MMS) according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25 µg of chromatin, 2 µg of ab208019 (red), and 20 µl of Protein A/G sepharose beads. 2μg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (SYBR green approach). rDNA primers used are located in the region of ribosomal gene loci following the publication PMID:21029860(rDNA 1: rDNA 13017F/13068R ; rDNA 2 : rDNA 1125F/1201R ; rDNA 3 : rDNA 30409F/30566R).

  • Immunohistochemical analysis of paraffin-embedded human breast tissue labeling Rad50 with ab208019 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining in human breast tissue (PMID: 15509680) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling Rad50 with ab208019 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining in rat testis is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling Rad50 with ab208019 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining in mouse testis (PMID: 10908350) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue labeling Rad50 with ab208019 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining in human breast carcinoma (PMID: 24642965) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • All lanes : Anti-Rad50 antibody [EPR20968] - ChIP Grade (ab208019) at 1/1000 dilution

    Lane 1 : K562 (human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
    Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 3 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
    Lane 4 : HT-29 (human colorectal adenocarcinoma cell line) whole cell lysate
    Lane 5 : NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate
    Lane 6 : ES-D3 (mouse embryonic multipotent stem Cell) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 153 kDa



    Blocking/Diluting buffer and concentration: 5% NFDM/TBST.

    Exposure time: Lanes 1-4: 8 seconds; Lanes 5-6: 24 seconds.

    The expression profile observed is consistent with what has been described in the literature (PMID: 26068589).

  • All lanes : Anti-Rad50 antibody [EPR20968] - ChIP Grade (ab208019) at 1/1000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

    Predicted band size: 153 kDa



    Blocking/Diluting buffer and concentration: 5% NFDM/TBST.

    Exposure time: Line 1: 3 minutes; Line 2: 41 seconds.

     

References

ab208019 has not yet been referenced specifically in any publications.

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