Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

Phospho-Rad18 (Ser403) Antibody #8393

Applications Reactivity Sensitivity MW (kDa) Source
W IP H (Mk) Endogenous 80, 90 Rabbit

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

Protocols

Specificity / Sensitivity

Phospho-Rad18 (Ser403) Antibody recognizes endogenous levels of Rad18 protein only when phosphorylated at Ser403. In some cell types, the antibody cross-reacts with a >200 kDa protein of unknown origin.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser403 of human Rad18 protein. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 or HeLa cells, untreated or UV-treated, using Phospho-Rad18 (Ser403) Antibody (upper) or a total Rad18 antibody (lower).

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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