HBV DNA and Protein Analysis

HBV core and PF DNAs were extracted as previously described and analyzed by Southern blotting [11 (link)]. Whole-cell extracts from a portion of transfected cells were fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and analyzed by western blotting for Tdp2 using polyclonal rabbit anti-human Tdp2 [20 (link)] or a commercial anti-Tdp2 antibody (Bethyl Labs) and HBV core protein using a mouse monoclonal antibody specific for the N-terminal end of core protein [41 (link)] respectively. To control for loading, western blotting for flap endonuclease 1 (FEN-1) or β-actin was carried out using monoclonal mouse anti-FEN-1 (BD Transduction Laboratories) or β-actin (Cell Signaling Technology). Southern and western blotting data were quantified by phosphorimaging and densitometry.

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Cui X., McAllister R., Boregowda R., Sohn J.A., Ledesma F.C., Caldecott K.W., Seeger C, & Hu J. (2015). Does Tyrosyl DNA Phosphodiesterase-2 Play a Role in Hepatitis B Virus Genome Repair?. PLoS ONE, 10(6), e0128401.

Publication 2015

anti-Tdp2 antibody β-actin

Actin Anti antibody Cell Cells extracts Densitometry Dnas Flap endonuclease 1 Human Monoclonal antibody Mouse Protein Protein n Rabbit Sds page

Corresponding Organization : University of Sussex

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Variable analysis

independent variables

HBV core and PF DNAs

dependent variables

HBV core protein

control variables

Loading control: FEN-1 or β-actin

controls

Positive control: Not specified
Negative control: Not specified

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