Lane 1 : Anti-pan Cadherin antibody [mAbcam22744] (ab22744) at 1 µg/ml (Blocked in 5% BSA)
Lane 2 : Anti-pan Cadherin antibody [mAbcam22744] (ab22744) at 1 µg/ml (Blocked in 5% Milk)

All lanes : Heart (Rat) Tissue Lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) (ab65485) at 1/3000 dilution

Performed under reducing conditions.

Predicted band size: 125 kDa


Exposure time: 30 seconds

ICC/IF image of ab22744 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab22744, 5µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Image-iT^TM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 phalloidin was used to label F-actin (red).

Overlay histogram showing HEK293 cells stained with ab22744 (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 (ab22744, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

All lanes : Anti-pan Cadherin antibody [mAbcam22744] (ab22744) at 1 µg/ml

Lane 1 : Heart (Rat) Tissue Lysate (blocked with 5% Milk)
Lane 2 : Heart (Mouse) Tissue Lysate (blocked with 5% Milk)
Lane 3 : Heart (Human) Tissue Lysate - adult normal tissue (blocked with 5% Milk)
Lane 4 : Heart (Rat) Tissue Lysate (blocked with 3% Milk)
Lane 5 : Heart (Mouse) Tissue Lysate (blocked with 3% Milk)
Lane 6 : Heart (Human) Tissue Lysate - adult normal tissue (blocked with 3% Milk)

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 125 kDa
Observed band size: 125 kDa
Additional bands at: 25 kDa, 58 kDa. We are unsure as to the identity of these extra bands.


Exposure time: 8 minutes

ab22744 staining pan Cadherin in mixed glia prepared from mouse brain by Immunocytochemistry/ Immunofluorescence. The cells were fixed in methanol, permeabilised in 0.5% (w/v) saponin and then blocked using 10% serum for 2 hours at 23°C. Samples were then incubated with primary antibody at 1/100 for 2 hours at 23°C. The secondary antibody used was a goat anti-mouse IgG conjugated to Alexa Fluor® 488 (green) used at a 1/400 dilution. Counterstained with DAPI (blue).

See Abreview

Anti-pan Cadherin antibody [mAbcam22744] (ab22744) at 1/500 dilution + Mouse cultured cortical neurons at 20 µg

Secondary
HRP-conjugated Goat Anti-Mouse IgG (H+L) polyclonal at 1/10000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 125 kDa


Exposure time: 1 minute


Blocking performed with 5% milk for 1 hour.
Primary diluted with PBS + 0.5% Tween20 and incubated for 12 hours at 4°C
Performed under denaturing conditions.

See Abreview

ICC/IF image of ab22744 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab22744, 5µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Image-iT^TM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 phalloidin was used to label F-actin (red).