Overview

  • Product name
  • Description
    Rabbit polyclonal to RNF8
  • Tested applications
    Suitable for: WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    A 14 amino acid peptide near the C terminus of Human RNF8 (NP_003949).

  • Positive control
    • WB: Human lung tissue lysate ICC/IF: Rat lung tissue

Properties

Applications

Our Abpromise guarantee covers the use of ab105362 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 Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 56 kDa.
ICC/IF Use a concentration of 20 µg/ml.

Target

  • Function
    E3 ubiquitin-protein ligase required for assembly of repair proteins to sites of DNA damage. Catalyzes the 'Lys-63'-linked ubiquitination of histone H2A and H2AX. Following DNA double-strand breaks (DSBs), it is recruited to the sites of damage by ATM-phosphorylated MDC1, mediates the ubiquitination of histones H2A and H2AX, thereby promoting the formation of TP53BP1 and BRCA1 ionizing radiation-induced foci (IRIF). Promotes the formation of 'Lys-63'-linked polyubiquitin chains and functions with the specific ubiquitin-conjugating UBE2N/UBC13. Substrates that are polyubiquitinated at 'Lys-63' are usually not targeted for degradation. Enforces the G2/M DNA damage checkpoint. Controls the recruitment of UIMC1-BRCC3 (RAP80-BRCC36) and PAXIP1/PTIP to DNA damage sites following DNA double-strand breaks (DSBs). Ubiquitination of histone H2A requires UBE2N but not MMS2 (UBE2V2). May also ubiquitinate histone H2B. Catalyzes the 'Lys-63'-linked ubiquitination of PCNA. May be required for proper exit from mitosis after spindle checkpoint activation and may regulate cytokinesis. May play a role in the regulation of RXRA-mediated transcriptional activity. Not involved in RXRA ubiquitination by UBE2E2.
  • Tissue specificity
    Ubiquitous. In fetal tissues, highest expression in brain, thymus and liver. In adult tissues, highest levels in brain and testis, lowest levels in peripheral blood cells.
  • Pathway
    Protein modification; protein ubiquitination.
  • Sequence similarities
    Belongs to the RNF8 family.
    Contains 1 FHA domain.
    Contains 1 RING-type zinc finger.
  • Developmental stage
    Low levels at the G1-S boundary increase in intensity during S phase and until the end of the G2 phase. Abruptly decreases in late mitosis (at protein level). Barely detectable in anaphase.
  • Domain
    The FHA domain specifically recognizes and binds ATM-phosphorylated MDC1 and Thr-4827 phosphorylated HERC2.
  • Post-translational
    modifications
    Autoubiquitinated through 'Lys-48' and 'Lys-63' of ubiquitin. 'Lys-63' polyubiquitination is mediated by UBE2N. 'Lys-29'-type polyubiquitination is also observed, but it doesn't require its own functional RING-type zinc finger.
  • Cellular localization
    Nucleus. Midbody. Following DNA double-strand breaks, recruited to the sites of damage. During prophase, concomitant with nuclear envelope breakdown, localizes throughout the cell, with a dotted pattern. In telophase, again in the nucleus and also with a discrete dotted pattern in the cytoplasm. In late telophase and during cytokinesis, localizes in the midbody of the tubulin bridge joining the daughter cells. Does not seem to be associated with condensed chromosomes at any time during the cell cycle.
  • Information by UniProt
  • Database links
  • Alternative names
    • C3HC4 type zinc finger protein antibody
    • E3 ubiquitin protein ligase RNF8 antibody
    • E3 ubiquitin-protein ligase RNF8 antibody
    • FLJ12013 antibody
    • KIAA0646 antibody
    • Ring finger protein (C3HC4 type) 8 antibody
    • RING finger protein 8 antibody
    • Ring finger protein 8, E3 ubiquitin protein ligase antibody
    • RNF 8 antibody
    • RNF8 antibody
    • RNF8_HUMAN antibody
    • UBC13/UEV interacting ring finger protein antibody
    see all

Images

  • Lane 1 : Anti-RNF8 antibody (ab105362) at 1 µg/ml
    Lane 2 : Anti-RNF8 antibody (ab105362) at 2 µg/ml

    Lane 1 : Human lung tissue lysate
    Lane 2 : Human lung tissue lysate

    Lysates/proteins at 15 µg per lane.


    Predicted band size : 56 kDa
  • Immunofluorescent analysis of RNF8 in Rat lung tissue using ab105362 at 20 µg/ml.
  • Immunofluorescence of RNF8 in Rat Lung cells using ab105362 at 20 ug/ml.

References

This product has been referenced in:
  • Patel N  et al. miR-15a/miR-16 down-regulates BMI1, impacting Ub-H2A mediated DNA repair and breast cancer cell sensitivity to doxorubicin. Sci Rep 7:4263 (2017). WB ; Human . Read more (PubMed: 28655885) »
  • Zhao MJ  et al. Adenovirus-mediated downregulation of the ubiquitin ligase RNF8 sensitizes bladder cancer to radiotherapy. Oncotarget 7:8956-67 (2016). WB . Read more (PubMed: 26788910) »

See all 2 Publications for this product

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (EC9706)
Permeabilization
Yes - 0.1% Triton X-100
Specification
EC9706
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jan 11 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (esophageal tumor cells&293T)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (10%SDS)
Loading amount
80 µg
Specification
esophageal tumor cells&293T
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Username

Abcam user community

Verified customer

Submitted Dec 31 2015

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

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Verified customer

Submitted Nov 20 2014

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

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