Anti-Cytokeratin 19 antibody [EPR1579Y]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 19 antibody [EPR1579Y] (AB76539)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung cancer tissue sections labeling Cytokeratin 19 with Purified ab76539 at 1 : 1000 dilution (0.12 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1 : 0 dilution. PBS instead of the primary antibody was used as the negative control.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Cytokeratin 19 antibody [EPR1579Y] (AB76539)

Overlay histogram showing MCF7 cells stained with unpurified ab76539 (red line). The cells were fixed with 4% paraformaldehyde (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 (ab76539, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 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). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Cytokeratin 19 antibody [EPR1579Y] (AB76539)

Intracellular Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Cytokeratin 19 with purified ab76539 at 1/20 dilution (10 μg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor^® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• ICC

Lab

Immunocytochemistry - Anti-Cytokeratin 19 antibody [EPR1579Y] (AB76539)

ab76539 staining Cytokeratin 19 in wild-type MCF7 cells (top panel) and KRT19 knockout MCF7 cells (bottom panel). The cells were fixed with 100% methanol (5 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 with ab76539 at 1/500 dilution and ab7291 (Mouse monoclonal to alpha 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 rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) 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).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 19 antibody [EPR1579Y] (AB76539)

Immunohistochemical staining of human stomach adenocarcinoma tissue with ab76539 (unpurified) at1/100-1/250.

Heat mediated antigen retrieval was performed via the pressure cooker method before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 19 antibody [EPR1579Y] (AB76539)

Fluorescent immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using ab76539 (unpurified). Green- CK19 red-PI.

Heat mediated antigen retrieval was performed via the pressure cooker method before commencing with IHC staining protocol.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 19 antibody [EPR1579Y] (AB76539)

ab76539 (unpurified) stained HCT116 cells. The cells were 4% formaldehyde fixed for 10 minutes at room temperature and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab76539 at 1/200 dilution) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor^® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43 μM for 1hour at room temperature.

• IP

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Immunoprecipitation - Anti-Cytokeratin 19 antibody [EPR1579Y] (AB76539)

ab76539 (purified) at 1 : 20 dilution (2ug) immunoprecipitating Cytokeratin 19 in SKBR-3 whole cell lysate.
Lane 1 (input) : SK-BR-3 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10 μg
Lane 2 (+) : ab76539 & SKBR-3 whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab76539 in SKBR-3 whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1 : 1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-Cytokeratin 19 antibody [EPR1579Y] (ab76539)

Predicted band size: 44 kDa

false

• WB

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Western blot - Anti-Cytokeratin 19 antibody [EPR1579Y] (AB76539)

All lanes:

Western blot - Anti-Cytokeratin 19 antibody [EPR1579Y] (ab76539) at 1/10000 dilution

Lane 1:

T47D cell lysate at 10 µg

Lane 2:

HepG2 cell lysate at 10 µg

Lane 3:

SKBR-3 cell lysate at 10 µg

Secondary

All lanes:

HRP-labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 44 kDa

false