Anti-EGFR antibody [EPR39Y] - BSA and Azide free

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-EGFR antibody [EPR39Y] - BSA and Azide free (AB239877)

ab76153 at 1/100 dilution staining EGFR in A431 cells by Immunofluorescence.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76153).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-EGFR antibody [EPR39Y] - BSA and Azide free (AB239877)

Overlay histogram showing A431 cells stained with ab76153 (red line). The cells were fixed with 80% methanol (5 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 (ab76153, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76153).

• WB

Lab

Western blot - Anti-EGFR antibody [EPR39Y] - BSA and Azide free (AB239877)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76153).

Western blot : Anti-EGFR antibody [EPR39Y] ab76153 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin ab7291 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 160 kDa in Wild-type A549 cell lysates with no signal observed at this size in EGFR knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-EGFR antibody [EPR39Y] (<a href='/ko/products/primary-antibodies/egfr-antibody-epr39y-ab76153'>ab76153</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysates at 20 µg

Lane 2:

EGFR knockout A549 cell lysates at 20 µg

Lane 3:

Wild-type HeLa ab255928 cell lysates at 20 µg

Lane 4:

Western blot - Human EGFR knockout HeLa cell line (<a href='/ko/products/cell-lines/human-egfr-knockout-hela-cell-line-ab255385'>ab255385</a>) at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 134 kDa

Observed band size: 160 kDa

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