Alexa Fluor® 647 Anti-EGFR (phospho Y1068) antibody [EP774Y]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 647 Anti-EGFR (phospho Y1068) antibody [EP774Y] (AB205828)

Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling EGFR (phospho Y1068) with ab205828 at 1/100 dilution. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 40 mins. Positive staining on human breast without alkaline phosphatase treatment (image A). No signal was detected when tissues were treated with alkaline phosphatase (image B). The section was incubated with ab205828 for 60 mins at room temperature (shown in red). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 647 Anti-EGFR (phospho Y1068) antibody [EP774Y] (AB205828)

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling EGFR (phospho Y1068) with ab205828 at 1/100 dilution. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 40 mins. Positive staining on human breast cancer without alkaline phosphatase treatment (image A). No signal was detected when tissues were treated with alkaline phosphatase (image B). The section was incubated with ab205828 for 60 mins at room temperature (shown in red). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-EGFR (phospho Y1068) antibody [EP774Y] (AB205828)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized C2C12 (mouse myoblast) cells labelling EGFR (phospho Y1068) with ab205828 at 1/50 (10.0 μg/ml) dilution (Green). ab195887 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 488) was used to counterstain tubulin at 1/200 (2.5 μg/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue). Confocal image showing positive staining in C2C12 cells treated with EGF (10ng/ml, 30min) and no staining on the cells without EGF treatment. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-EGFR (phospho Y1068) antibody [EP774Y] (AB205828)

Flow Cytometry analysis of A431 (human epidermoid carcinoma) cells labeling EGFR (phospho Y1068) with purified ab205828 at 1/500 dilution(10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 647) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-EGFR (phospho Y1068) antibody [EP774Y] (AB205828)

ab205828 staining EGFR (phospho Y1092) in A431 cells +/-EGF (100ng/ml, 5min). The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h.

The cells were then incubated overnight at +4°C with ab205828 at 1 : 100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).