Immunohistochemical analysis of human breast cancer tissue labeling E Cadherin with ab1416.

Anti-E Cadherin antibody [HECD-1] - Intercellular Junction Marker (ab1416) at 1/50 dilution + LNCaP (human prostate cancer cell line) whole cell lyate

See Abreview

All lanes : Anti-E Cadherin antibody [HECD-1] - Intercellular Junction Marker (ab1416) at 1/1000 dilution

All lanes : MDA-MB-468 cells whole cell lysate

Lysates/proteins at 80 µg/ml per lane.

Secondary
All lanes : HRP-conjugated polyclonal donkey anti-mouse IgG

Performed under reducing conditions.

Observed band size: 110 kDa why is the actual band size different from the predicted?


Exposure time: 30 seconds

See Abreview

Lanes 1-2 : Anti-E Cadherin antibody [HECD-1] - Intercellular Junction Marker (ab1416) at 1/50 dilution
Lanes 3-4 : Anti-GAPDH at 1 µg/ml

Lanes 1 & 3 : MDA-MB-231 cell lysate
Lanes 2 & 4 : MCF-7 cell lysate

Blocking buffer: 3% milk

Overlay histogram showing MCF7 cells stained with ab1416 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab1416, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-mouse IgG (H+L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 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. Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

Flow cytometric analysis of A431 (human epidermoid carcinoma cell line) cell line labeling E Cadherin with ab1416 at 1/100 dilution (2) compared with an isotype control (1).

See Abreview

ab1416 staining E Cadherin in human stomach tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent formaldehyde fixation before heat mediated antigen retrieval in Citrate pH 6.0 and then blocking with 5% serum for 1 hour at 23°C was performed. The primary antibody was diluted 1/100 and incubated with sample for 1 hour at 23°C. A HRP conjugated goat polyclonal to mouse IgG was used undiluted as secondary antibody.

See Abreview

Immunohistochemistry of human breast carcinoma staining E Cadherin with ab1416 at 5μg/ml