| Products/Services Used | Details | Operation |
|---|---|---|
| Protein Electrophoresis and Western | 3xFLAG-tagged proteins were immunoprecipitated overnight at 4°C using 25 μl GenScript Anti-FLAG G1 Affinity Resin according to the manufacturer’s protocol and washed resins were resuspended in TBS.Proteins were electrophoresed in a 4%–20% or 4%–12% SurePAGE gel and Tris–MOPS–SDS running buffer (GenScript).The membrane was blocked (PBS, 5% non-fat milk) and probed with rabbit anti-DYDDDDK primary antibody (GenScript, 1:2500; PBS–0.05% Tween 20, 1% non-fat milk) and rabbit IgG (HRP) secondary antibody (Abcam, 1:10 000; PBS–0.05% Tween 20, 1% non-fat milk) with washes after antibody incubations (Phosphate buffered saline–0.05% Tween 20). | Get A Quote |
DNA polymerase theta (Pol θ) is an error-prone translesion polymerase that becomes crucial for DNA double-strand break repair when cells are deficient in homologous recombination or non-homologous end joining. In some organisms, Pol θ also promotes tolerance of DNA interstrand crosslinks. Due to its importance in DNA damage tolerance, Pol θ is an emerging target for treatment of cancer and disease. Prior work has characterized the functions of the Pol θ helicase-like and polymerase domains, but the roles of the linker domain are largely unknown. Here, we show that the Drosophila melanogaster Pol θ linker domain promotes proper egg development and is required for repair of DNA double-strand breaks and inter... More
