Revision 1
#14978
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For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:
W, IP
Reactivity:
H
Sensitivity:
Endogenous
MW (kDa):
80, 90
Source/Isotype:
Rabbit IgG
UniProt ID:
#Q9NS91
Entrez-Gene Id:
56852
Product Usage Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Immunoprecipitation | 1:50 |
Storage
Specificity/Sensitivity
Phospho-Rad18 (Ser403) (D2T6W) Rabbit mAb recognizes endogenous levels of Rad18 protein only when phosphorylated at Ser403.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser403 of human Rad18 protein.
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).
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
W: Western Blotting IP: Immunoprecipitation
Cross-Reactivity Key
H: Human M: Mouse R: Rat Hm: Hamster Mk: Monkey Vir: Virus Mi: Mink C: Chicken Dm: D. melanogaster X: Xenopus Z: Zebrafish B: Bovine Dg: Dog Pg: Pig Sc: S. cerevisiae Ce: C. elegans Hr: Horse GP: Guinea Pig Rab: Rabbit G: Goat All: All Species Expected
Trademarks and Patents
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.
Limited Uses
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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.
Revision 1
Western blot analysis of extracts from 293 cells, untreated (-) or UV-treated (+), using Phospho-Rad18 (Ser403) (D2T6W) Rabbit mAb (upper) and Rad18 (D2B8) XP® Rabbit mAb #9040 (lower).
Western blot analysis of extracts from 293 cells, untreated (-) or UV-treated (+), using Phospho-Rad18 (Ser403) (D2T6W) Rabbit mAb. Following transfer, blots were either mock treated (left) or treated with calf intestinal phosphatase (CIP, right).
Immunoprecipitation of phospho-Rad18 (Ser403) from UV-treated 293 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Phospho-Rad18 (Ser403) (D2T6W) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot was performed using Phospho-Rad18 (Ser403) (D2T6W) Rabbit mAb.