anti human polλ antibody  (Novus Biologicals)

Journal: Life Science Alliance

Article Title: A novel role of DNA polymerase λ in translesion synthesis in conjunction with DNA polymerase ζ

doi: 10.26508/lsa.202000900

Figure Lengend Snippet: (A) Western blot analyses of the efficiency of siRNA knockdown of human Polλ in GM637 HFs (left) or of mouse Polλ in MEFs (right). NC: negative control siRNA. β-tubulin was used as the loading control (bottom). (B) Western blot analysis of Polλ expression in GM637 HFs (left) or in Polλ −/− MEFs (right) expressing 3xFlag-siRNA resistant wild-type human Polλ, 3xFlag-siRNA resistant catalytic mutant (D427A D429A) of human Polλ, or the vector control. Human Polλ has been reported to be phosphorylated at multiple sites in vivo (Wimmer U, Ferrari E, Hunziker P, Hübscher U (2008) EMBO Rep 9: 1027–1033). The distinct upper band in the right panel likely is phosphorylated Polλ. (C) Western blot analyses of expression of 3× Flag-siRNA resistant WT Polλ, 3× Flag-siRNA resistant N-terminally deleted (245–575) Polλ, or the vector control in GM637 HFs. GM637 HFs or MEFs were treated with Polλ siRNA for 48 h. (A, B, C) Polλ expression was determined by Western blot analysis with Polλ ab (A) or Flag ab (B, C). βtubulin or lamin B1 were used as the loading control. (D) Western blot analyses of expression of 3× Flag-siRNA resistant WT Rev3, 3× Flag-siRNA resistant catalytic mutant (D2781A and D2783A) Rev3, or the vector control in GM637 HFs. GM637 HFs were treated with Rev3 siRNA for 48 h. Rev3 expression was determined by Western blot analysis with Flag ab. * indicates a non-specific band. β-tubulin or lamin B1 were used as the loading control.

Article Snippet: The membranes were probed with antibodies against human Polλ (Bethyl Lab) or Flag (Sigma-Aldrich), followed by appropriate secondary antibodies conjugated with horseradish peroxidase.

Techniques: Western Blot, Knockdown, Negative Control, Control, Expressing, Mutagenesis, Plasmid Preparation, In Vivo