Alexa Fluor® 647 Anti-EGFR antibody [EP38Y]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-EGFR antibody [EP38Y] (AB192982)

ab192982 staining EGFR in HeLa cells. The cells were fixed with 100% methanol (5 min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Triton X-100 for 1hr. The cells were then incubated with ab192982 at a working dilution of 1 in 50 (shown in red) and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin) at 1μg/ml overnight at +4°C, followed by a further incubation at room temperature for 1hr with AlexaFluor^® 488 Goat anti-mouse IgG (H&L - preadsorbed) (ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

Image was taken with a Confocal microscope (Leica-microsystems, TCS SP8).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-EGFR antibody [EP38Y] (AB192982)

Overlay histogram showing HeLa cells stained with ab192982 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab192982, 2μl/1x10⁶ cells) for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (Alexa Fluor^® 647) (1μg/1x10⁶ cells) for 30 min at 22°C. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a 25mW red solid state diode laser (635nm) and 675/30 bandpass filter.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-EGFR antibody [EP38Y] (AB192982)

ab192982 staining EGFR in A431 cells. 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 ab192982 at 1/1000 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).

This product also gave a positive signal under the same testing conditions in A431 cells fixed with 100% methanol (5 min).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 647 Anti-EGFR antibody [EP38Y] (AB192982)

Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling EGFR with ab192982 at 1/50 dilution.

Positive staining on human lung cancer.

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

Heat mediated antigen retrieval was performed 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® 647 Anti-EGFR antibody [EP38Y] (AB192982)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling EGFR with ab192982 at 1/50 dilution

Positive staining on squamous epithelial of human tonsil.

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

Heat mediated antigen retrieval was performed 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® 647 Anti-EGFR antibody [EP38Y] (AB192982)

Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling EGFR with ab192982 at 1/50 dilution.

Positive staining on human placenta.

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

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 40 mins.