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Western blot using EGFR (phospho Y1197) polyclonal antibody (Cat # PAB10009) shows detection of a band at ~170 KDa corresponding to human EGFR (arrowhead).
Staining is not seen in unstimulated A-431 cells (Lane 1), but is seen when A-431 cells are stimulated with EGF (50 ng/mL for 15 min) (lane 2). Approximately 30 ug of lysate was separated on a 4-20% Tris-Glycine gel by SDS-PAGE and transferred onto nitrocellulose.
After blocking the membrane was probed with the primary antibody diluted to 1 : 250.
Reaction occurred overnight at 4°C followed by washes and reaction with a 1 : 10,000 dilution of IRDye™800 conjugated Gt-a-Rabbit IgG [H&L] MX for 45 min at room temperature (800 nm channel, green).
Molecular weight estimation was made by comparison to prestained MW markers in lane M (700 nm channel, red).
IRDye™800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR.
IRDye is a trademark of LI-COR, Inc.

Immunohistochemical staining of EGFR (phospho Y1197) polyclonal antibody (Cat # PAB10009) was used at 5 ug/mL to detect signal in a variety of tissues including multi-human, multi-brain and multi-cancer slides.
This image shows faintly to moderately positive staining of placental trophoblasts at 40X. Tissue was formalin-fixed and paraffin embedded. The image shows localization of the antibody as the precipitated red signal, with a hematoxylin purple nuclear counterstain.
Personal Communi-cation, Tina Roush, Life Span Biosciences, Seattle, WA.