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Western blot All lanes: RNF8 antibody at 11microg/ml Lane 1: Hela whole cell lysate Lane 2: 293T whole cell lysate Secondary Goat polyclonal to rabbit IgG at 1/10000 dilution Predicted band size: 56, 11, 51 kDa Observed band size: 56 kDa
Western blot All lanes: RNF8 antibody at 11microg/ml Lane 1: Hela whole cell lysate Lane 2: 293T whole cell lysate Secondary Goat polyclonal to rabbit IgG at 1/10000 dilution Predicted band size: 56, 11, 51 kDa Observed band size: 56 kDa
Western blot All lanes: RNF8 antibody at 11microg/ml Lane 1: Hela whole cell lysate Lane 2: 293T whole cell lysate Secondary Goat polyclonal to rabbit IgG at 1/10000 dilution Predicted band size: 56, 11, 51 kDa Observed band size: 56 kDa

RNF8 Antibody

CSB-PA019898ESR2HU
Cusabio
ApplicationsWestern Blot, ELISA, ImmunoHistoChemistry
Product group Antibodies
TargetRNF8
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Overview

  • Supplier
    Cusabio
  • Product Name
    RNF8 Antibody
  • Delivery Days Customer
    20
  • Applications
    Western Blot, ELISA, ImmunoHistoChemistry
  • Certification
    Research Use Only
  • Clonality
    Polyclonal
  • Conjugate
    Unconjugated
  • Gene ID9025
  • Target name
    RNF8
  • Target description
    ring finger protein 8
  • Target synonyms
    C3HC4-type zinc finger protein; E3 ubiquitin-protein ligase RNF8; hRNF8; ring finger protein (C3HC4 type) 8; ring finger protein 8, E3 ubiquitin protein ligase; RING-type E3 ubiquitin transferase RNF8; UBC13/UEV-interacting ring finger protein
  • Host
    Rabbit
  • Isotype
    IgG
  • Protein IDO76064
  • Protein Name
    E3 ubiquitin-protein ligase RNF8
  • Scientific Description
    E3 ubiquitin-protein ligase that plays a key role in DNA damage signaling via 2 distinct roles: by mediating the Lys-63-linked ubiquitination of histones H2A and H2AX and promoting the recruitment of DNA repair proteins at double-strand breaks (DSBs) sites, and by catalyzing Lys-48-linked ubiquitination to remove target proteins from DNA damage sites. Following DNA DSBs, it is recruited to the sites of damage by ATM-phosphorylated MDC1 and catalyzes the Lys-63-linked ubiquitination of histones H2A and H2AX, thereby promoting the formation of TP53BP1 and BRCA1 ionizing radiation-induced foci (IRIF). Also controls the recruitment of UIMC1-BRCC3 (RAP80-BRCC36) and PAXIP1/PTIP to DNA damage sites. Also recruited at DNA interstrand cross-links (ICLs) sites and catalyzes Lys-63-linked ubiquitination of histones H2A and H2AX, leading to recruitment of FAAP20/C1orf86 and Fanconi anemia (FA) complex, followed by interstrand cross-link repair. H2A ubiquitination also mediates the ATM-dependent transcriptional silencing at regions flanking DSBs in cis, a mechanism to avoid collision between transcription and repair intermediates. Promotes the formation of Lys-63-linked polyubiquitin chains via interactions with the specific ubiquitin-conjugating UBE2N/UBC13 and ubiquitinates non-histone substrates such as PCNA. Substrates that are polyubiquitinated at Lys-63 are usually not targeted for degradation. Also catalyzes the formation of Lys-48-linked polyubiquitin chains via interaction with the ubiquitin-conjugating UBE2L6/UBCH8, leading to degradation of substrate proteins such as CHEK2, JMJD2A/KDM4A and KU80/XRCC5: it is still unclear how the preference toward Lys-48- versus Lys-63-linked ubiquitination is regulated but it could be due to RNF8 ability to interact with specific E2 specific ligases. For instance, interaction with phosphorylated HERC2 promotes the association between RNF8 and UBE2N/UBC13 and favors the specific formation of Lys-63-linked ubiquitin chains. Promotes non-homologous end joining (NHEJ) by promoting the Lys-48-linked ubiquitination and degradation the of KU80/XRCC5. Following DNA damage, mediates the ubiquitination and degradation of JMJD2A/KDM4A in collaboration with RNF168, leading to unmask H4K20me2 mark and promote the recruitment of TP53BP1 at DNA damage sites. In addition to its function in damage signaling, also plays a role in higher-order chromatin structure by mediating extensive chromatin decondensation. Involved in the activation of ATM by promoting histone H2B ubiquitination, which indirectly triggers histone H4 Lys-16 acetylation (H4K16ac), establishing a chromatin environment that promotes efficient activation of ATM kinase. Required in the testis, where it plays a role in the replacement of histones during spermatogenesis. At uncapped telomeres, promotes the joining of deprotected chromosome ends by inducing H2A ubiquitination and TP53BP1 recruitment, suggesting that it may enhance cancer development by aggravating telomere-induced genome instability in case of telomeric crisis. Promotes the assembly of RAD51 at DNA DSBs in the absence of BRCA1 and TP53BP1 Also involved in class switch recombination in immune system, via its role in regulation of DSBs repair. 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.
  • Storage Instruction
    -20°C or -80°C
  • UNSPSC
    12352203

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Figure 1. Western blot analysis of RNF8 using anti-RNF8 antibody (A00707-2). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human Hela whole cell lysates, Lane 2: human HEK293 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-RNF8 antigen affinity purified polyclonal antibody (Catalog # A00707-2) at 0.5 microg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for RNF8 at approximately 56 kDa. The expected band size for RNF8 is at 56 kDa.
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