phosphorylated c src  (Proteintech)

Journal: PLoS ONE

Article Title: EGF Receptor Exposed to Oxidative Stress Acquires Abnormal Phosphorylation and Aberrant Activated Conformation That Impairs Canonical Dimerization

doi: 10.1371/journal.pone.0023240

Figure Lengend Snippet: Serum-starved A549 cells were incubated (or not) with 1 µM AG1478 or 5 µM PP1 for 30 min. Then, the cells were treated for 30 min. with 100 ng/ml EGF or 1 U/ml GO, as indicated. EGFR was IPed from cell lysates, resolved by SDS-PAGE and IBed for total receptor, total tyrosine phosphorylation (p-EGFR) and specific Tyr-residue phosphorylation level (Y845, Y1068, Y1086, and Y1173). Protein aliquots of the cell lysates were also directly IBed for total and Y416 phosphorylated (active) c-Src (p-Src).

Article Snippet: Primary Abs used in this study for the IBs were: α2232 (αEGFR, Cell Signaling, 1∶1000), αc-Src (Santa Cruz Biotech, 1∶2000), α tyrosine Y416 phosphorylated (p-) c-Src (Cell Signaling, 1∶2000), αp-Y20 (Santa Cruz Biotech, 1∶3000), αp-Y1173 EGFR (Santa Cruz Biotech, 1∶1000), αp-Y1086, αp-Y1068 and αp-Y845 EGFR (Cell Signaling, 1∶1000).

Techniques: Incubation, SDS Page

Journal: PLoS ONE

Article Title: EGF Receptor Exposed to Oxidative Stress Acquires Abnormal Phosphorylation and Aberrant Activated Conformation That Impairs Canonical Dimerization

doi: 10.1371/journal.pone.0023240

Figure Lengend Snippet: A549 cells were incubated (or not) with 5 µM PP1 for 45 min. and then treated (or not) for 15 min. with 100 ng/ml EGF or 30 min. 1 U/ml GO. A . EGFR was IPed from total cell lysates with the mAb 528 and IBed for Y416 phosphorylated c-Src (p-Src) and for total EGFR, as indicated. B . c-Src was IPed from total cell lysates and IBed for total c-Src and EGFR, as indicated.

Article Snippet: Primary Abs used in this study for the IBs were: α2232 (αEGFR, Cell Signaling, 1∶1000), αc-Src (Santa Cruz Biotech, 1∶2000), α tyrosine Y416 phosphorylated (p-) c-Src (Cell Signaling, 1∶2000), αp-Y20 (Santa Cruz Biotech, 1∶3000), αp-Y1173 EGFR (Santa Cruz Biotech, 1∶1000), αp-Y1086, αp-Y1068 and αp-Y845 EGFR (Cell Signaling, 1∶1000).

Techniques: Incubation

Journal: PLoS ONE

Article Title: EGF Receptor Exposed to Oxidative Stress Acquires Abnormal Phosphorylation and Aberrant Activated Conformation That Impairs Canonical Dimerization

doi: 10.1371/journal.pone.0023240

Figure Lengend Snippet: A . Serum-starved A549 cells were treated, or not, with 100 ng/ml EGF for 15 min. or 2% (w/v) MβCD for 1 h and cell lysates were IBed for total and Tyr-phosphorylated EGFR. B and C . Cells were treated with EGF as in A or with 1 U/ml GO for 30 min. in the absence or presence of 2 mM MβCD-cholesterol complex (CC), prepared as described in “ ”. Cell lysates were separated by SDS-PAGE and IBed for total and Tyr-phosphorylated EGFR ( B ) or total and Y416 phosphorylated Src ( C ).

Article Snippet: Primary Abs used in this study for the IBs were: α2232 (αEGFR, Cell Signaling, 1∶1000), αc-Src (Santa Cruz Biotech, 1∶2000), α tyrosine Y416 phosphorylated (p-) c-Src (Cell Signaling, 1∶2000), αp-Y20 (Santa Cruz Biotech, 1∶3000), αp-Y1173 EGFR (Santa Cruz Biotech, 1∶1000), αp-Y1086, αp-Y1068 and αp-Y845 EGFR (Cell Signaling, 1∶1000).

Techniques: SDS Page

Journal: PLoS ONE

Article Title: EGF Receptor Exposed to Oxidative Stress Acquires Abnormal Phosphorylation and Aberrant Activated Conformation That Impairs Canonical Dimerization

doi: 10.1371/journal.pone.0023240

Figure Lengend Snippet: A549 cells were seeded on cover-glasses, serum starved for 1 h and treated (or not) with 1 U/ml GO as indicated. After treatments, ceramide, EGFR phosphorylated on Y1173 and Y416 phosphorylated c-Src were localized in situ by IF as indicated in “ ”; nuclei were stained by DAPI.

Article Snippet: Primary Abs used in this study for the IBs were: α2232 (αEGFR, Cell Signaling, 1∶1000), αc-Src (Santa Cruz Biotech, 1∶2000), α tyrosine Y416 phosphorylated (p-) c-Src (Cell Signaling, 1∶2000), αp-Y20 (Santa Cruz Biotech, 1∶3000), αp-Y1173 EGFR (Santa Cruz Biotech, 1∶1000), αp-Y1086, αp-Y1068 and αp-Y845 EGFR (Cell Signaling, 1∶1000).

Techniques: In Situ, Staining

Journal: PLoS ONE

Article Title: EGF Receptor Exposed to Oxidative Stress Acquires Abnormal Phosphorylation and Aberrant Activated Conformation That Impairs Canonical Dimerization

doi: 10.1371/journal.pone.0023240

Figure Lengend Snippet: A549 cells were seeded on cover-glasses, serum starved for 1 h and treated (or not) with 1 U/ml GO as indicated. After treatments, ceramide ( A and B ), EGFR phosphorylated on Y1173 ( A ) and Y416 phosphorylated c-Src ( B ) were localized in situ by IF as indicated in “ ”; nuclei were stained by DAPI. White arrows indicate regions where p-Y1173 EGFR and ceramide ( A ) or p-Y416 c-Src (p-Src) and ceramide ( B ) co-localized under ox-stress (GO). Z-stack sections of cells have been discriminated by confocal microscopy: the panels show the merge of all of the Z-stack sections.

Article Snippet: Primary Abs used in this study for the IBs were: α2232 (αEGFR, Cell Signaling, 1∶1000), αc-Src (Santa Cruz Biotech, 1∶2000), α tyrosine Y416 phosphorylated (p-) c-Src (Cell Signaling, 1∶2000), αp-Y20 (Santa Cruz Biotech, 1∶3000), αp-Y1173 EGFR (Santa Cruz Biotech, 1∶1000), αp-Y1086, αp-Y1068 and αp-Y845 EGFR (Cell Signaling, 1∶1000).

Techniques: In Situ, Staining, Confocal Microscopy