Anti hpv16 e7 antibody

Poly methyl methacrylate (PMMA) was purchased from McMaster-Carr (Atlanta, GA). Double-sided adhesive polymer film (DSA) was purchased from iTapestore (Scotch Plains, NJ). Millipore isopore polycarbonate membrane was obtained from Merck KGaA, (Darmstadt, Germany). Plastic tubing was purchased from Cole-Parmer (Vernon Hills, IL). Phosphate buffered saline (PBS) was obtained from Life Technologies (Grand Island, NY). Human IgG ELISA Kit was purchased from Abcam (ab100547) (Cambridge, MA). Whole Blood (EDTA) was obtained from Stanford Blood Center (Palo Alto, CA). 3,3′,5,5′-Tetramethylbenzidine (TMB substrate), and Bovine serum albumin powder (BSA) were purchased from Sigma Aldrich (St. Louis, MO). Peroxidase AffiniPure Sheep Anti-Mouse IgG (H + L) (HRP) and Peroxidase AffiniPure Goat Anti-Human IgG, Fcγ Fragment Specific was purchased from Jackson ImmunoResearch Laboratories, Inc. (West Grove, PA). BupH Carbonate-Bicarbonate buffer packs were obtained from Thermo Scientific (Rockford, IL). anti-HPV16 E7 antibody was purchased from Santa Cruz Biotechnology (sc-6981, Dallas, TX). HPV16 E7 protein was provided by Professor Karen Anderson, Arizona State University (Tempe, AZ).

For co-immunoprecipitation assays, total cell extracts were incubated with an anti-HPV16 E7 antibody (Santa Cruz Biotechnology, Cat. No. sc-6981) at 4 °C. Immune complexes were recovered using protein A-agarose beads (GenDepot). For GST-pull down assays, bacteria were lysed in lysis buffer (150 mM NaCl, 50 mM Tris-HCl, pH7.5, 10 mM EDTA, 3 mg/mL lysozyme, 1% Triton X-100, and protease inhibitors), and GST fusion proteins were purified with glutathione agarose beads (Takara Bio, Mountain View, CA, USA) according to the manufacturer’s instruction. The resulting complexes were then incubated with cell lysates.