Journal: PLoS ONE

Article Title: Measuring EGFR Separations on Cells with ∼10 nm Resolution via Fluorophore Localization Imaging with Photobleaching

doi: 10.1371/journal.pone.0062331

Figure Lengend Snippet: (A) Single-molecule TIRF image of glass-immobilised 13.3 nm DNA rulers, labelled at each end with Atto 647 N. (B) Single-molecule TIRF image of T47D cells labelled with anti-EGFR Affibody-Atto 647 N. Scale bar = 8 µm.

Article Snippet: Protein S resin (Sigma) was washed twice with PBS and incubated with anti-EGFR antibody (Cell Signaling Technology #2256) for 1 h at room temperature, then washed twice with PBS to remove unbound antibody.

Techniques:

Journal: PLoS ONE

Article Title: Measuring EGFR Separations on Cells with ∼10 nm Resolution via Fluorophore Localization Imaging with Photobleaching

doi: 10.1371/journal.pone.0062331

Figure Lengend Snippet: (A) Western blots of A431 cell immunoprecipitates (I) and whole cell lysates (C) from unstimulated cells (blank) and cells stimulated with either 100 nM EGF or 2 nM EGFR Affibody. Proteins were probed (left to right) for non-specific tyrosine phosphorylation of proteins (Anti-pY4G10), tyrosine phosphorylation of EGFR (Anti-pY1045 EGFR) to show activated receptor, and presence of EGFR (Anti-EGFR). (B) Results of densitometry normalized against the total EGFR and relative quantity expressed as a fold change compared to an unstimulated control.

Article Snippet: Protein S resin (Sigma) was washed twice with PBS and incubated with anti-EGFR antibody (Cell Signaling Technology #2256) for 1 h at room temperature, then washed twice with PBS to remove unbound antibody.

Techniques: Western Blot, Phospho-proteomics, Control