Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab15850 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.
WB Use a concentration of 0.1 - 1 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 56 kDa).Can be blocked with Human RNF8 peptide (ab23278).

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

  • Anti-RNF8 antibody (ab15850) at 0.1 µg/ml + Lysates prepared from human lung tissues at 35 µg

    Predicted band size : 56 kDa
  • ab15850 staining RNF8 in human U2OS cells by Immunocytochemistry/ Immunofluorescence.
    Cells were fixed in formaldehyde, permeabilized using NP40, blocked with 5% BSA fo 1 hour at 25°C and then incubated with ab15850 at a 1/100 dilution. The secondary used was an Alexa-Fluor 488 conjugated donkey anti-goat polyclonal used at a 1/500 dilution.

    See Abreview

References

ab15850 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (10%)
Sample
Human Cell lysate - whole cell (U2OS)
Specification
U2OS
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Oct 03 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (U2OS)
Specification
U2OS
Fixative
Formaldehyde
Permeabilization
Yes - NP40
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Andrew Cobb

Verified customer

Submitted Mar 06 2012

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

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