Alexa Fluor® 488 Anti-EGFR (phospho Y1068) antibody [EP774Y]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-EGFR (phospho Y1068) antibody [EP774Y] (AB205827)

Immunocytochemistry analysis of ab205827 staining EGFR (phospho Y1068) in A431 cells +/-EGF (100 ng/ml, 5min). The cells were fixed with 100% methanol (5 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 1 hour.

The cells were then incubated overnight at +4°C with ab205827 at 1 : 100 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

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

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Anti-EGFR (phospho Y1068) antibody [EP774Y] (AB205827)

Flow Cytometry analysis of A431 (human epidermoid carcinoma) cells labeling EGFR (phospho Y1068) with purified ab205827 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® 488) 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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 488 Anti-EGFR (phospho Y1068) antibody [EP774Y] (AB205827)

Immunohistochemical analysis of paraffin-embedded tissue labeling EGFR (phospho Y1068) with ab205827 at 1/50 dilution.

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 primary antibody was incubated for 60 mins at room temperature (shown in green). 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).

Counterstained with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 40 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 488 Anti-EGFR (phospho Y1068) antibody [EP774Y] (AB205827)

Immunohistochemical analysis of paraffin-embedded tissue labeling EGFR (phospho Y1068) with ab205827 at 1/50 dilution.

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 primary antibody was incubated for 60 mins at room temperature (shown in green). 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).

Counterstained with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 40 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 488 Anti-EGFR (phospho Y1068) antibody [EP774Y] (AB205827)

Immunohistochemical analysis of paraffin-embedded tissue labeling EGFR (phospho Y1068) with ab205827 at 1/50 dilution.

Negative control : no staining on human liver.

The primary antibody was incubated for 60 mins at room temperature (shown in green). 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).

Counterstained with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 40 mins.