Anti-EpCAM antibody [VU-1D9]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-EpCAM antibody [VU-1D9] (AB187372)

ab187372 staining EpCAM in wild-type A431 cells (top panel) and EpCAM knockout A431 cells (bottom panel). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% PBS-Tween 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 with ab187372 at 0.1μg/ml concentration and ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor^® 488) (ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor^® 594) (ab150080) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI. This antibody performed similarly using 100% methanol fixation. Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a single confocal section is shown.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-EpCAM antibody [VU-1D9] (AB187372)

ab187372 staining EpCAM in wild-type A431 cells (top panel) and EpCAM knockout A431 cells (bottom panel). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% PBS-Tween 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 with ab187372 at 0.5μg/ml concentration and ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor^® 488) (ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor^® 594) (ab150080) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-EpCAM antibody [VU-1D9] (AB187372)

ab187372 staining EpCAM in T47D positive cells (top panel) and HeLa negative cells (bottom panel). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% PBS-Tween 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 with ab187372 at 1μg/ml concentration and ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor^® 488) (ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor^® 594) (ab150080) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI. This antibody performed similarly using 100% methanol fixation. Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a single confocal section is shown.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-EpCAM antibody [VU-1D9] (AB187372)

ab187372 staining EpCAM in HT-29 cell line (top panel) and Jurkat negative cells (bottom panel). The cells were fixed with 4% paraformaldehyde then permeabilized with 0.1% TritonX-100. The cells were then incubated with ab187372 at 1/250 concentration and counterstained with ab179513 (Anti-beta Tubulin rabbit monoclonal antibody) at 1/200 dilution, followed by secondary antibody ab150113 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488)) at 1/1000 dilution (shown in green) and counterstained with ab150080 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594)) at 1/500 dilution (shown in red). Nuclear DNA was labelled in blue with DAPI.

Confocal image showing membranous staining in HT-29 cell line.
Negative control : Jurkat (PMID : 29352248）

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-EpCAM antibody [VU-1D9] (AB187372)

Flow cytometric analysis of Jurkat (Human T cell leukemia T lymphocyte, Left) / HT-29 (Human colorectal adenocarcinoma epithelial cell, Right) labeling EpCAM with ab187372 at 1/1000 dilution, followed by secondary antibody ab150113 (Goat anti mouse IgG (Alexa Fluor® 488)) at 1/2000 dilution (Red). Compared with a Mouse monoclonal IgG isotype control (Black) and an unlabelled control (Cell without incubation with primary antibody and secondary antibody) (Blue). Gated on viable cells.

Negative control : Jurkat (PMID : 29352248)