Cell Signaling Technology
XP Monoclonal Antibody

Product Pathways - DNA Damage

Rad18 (D2B8) XP® Rabbit mAb #9040

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC H Endogenous 80, 90 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

* Product-specific protocol.

Specificity / Sensitivity

Rad18 (D2B8) XP® Rabbit mAb recognizes endogenous levels of total Rad18 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human Rad18 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Rad18 (D2B8) XP® Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or UV-treated (right), showing translocation of Rad18 from large spontaneous foci to numerous small damage-induced foci, using Rad18 (D2B8) XP® Rabbit mAb (green). Actin filaments were labeled with β-Actin (8H10D10) Mouse mAb #3700 (red).

Background

DNA damage, if not repaired, can lead to genome instability and tumorigenesis. Eukaryotic cells use multiple (sometimes overlapping) signaling pathways to respond to agents that cause various types of DNA lesions. Downstream molecules in DNA repair pathways converge on the sites of DNA damage, resulting in cell cycle arrest and repair or apoptosis (1). Rad18 is an E3 ubiquitin ligase recruited to sites of DNA damage. Along with the E2 ubiquitin ligase Rad6, Rad18 is responsible for monoubiquitination of DNA damage proteins including the replication clamp PCNA and the Fanconi anemia core protein FANCD2. Monoubiquitination of these proteins signals to downstream effector molecules and results in the repair of either post-replication repair lesions via the translesion synthesis (TLS) pathway or DNA double strand breaks via homologous recombination (2-4). Phospho-proteomic studies indicate that Ser403 of Rad18 may be phosphorylated by ATM/ATR in response to DNA damage-inducing agents (5,6).

  1. Helleday, T. et al. (2008) Nat Rev Cancer 8, 193-204.
  2. Huang, J. et al. (2009) Nat Cell Biol 11, 592-603.
  3. Song, I.Y. et al. (2010) J Biol Chem 285, 31525-36.
  4. Ting, L. et al. (2010) DNA Repair (Amst) 9, 1241-8.
  5. Mu, J.J. et al. (2007) J Biol Chem 282, 17330-4.
  6. Matsuoka, S. et al. (2007) Science 316, 1160-6.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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