Anti-Cytokeratin 19 antibody [LP2K]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 19 antibody [LP2K] (AB254186)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue labeling Cytokeratin 19 with ab254186 at 1/10000 (0.105 μg/ml) dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous staining on human lung carcinoma is observed. The section was incubated with ab254186 for 30 mins at room temperature and followed by specific mouse IgG antibody (ab190482) for 8mins. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 19 antibody [LP2K] (AB254186)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 100% Methanol permeabilized MCF7 (human breast adenocarcinoma epithelial cell) cells labelling Cytokeratin 19 with ab254186 at 1/500 (2.096 μg/ml) dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in MCF7 cells.

ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at a 1/1000 dilution (Red). The nuclear counterstain was DAPI (Blue).

Negative control 1 : ab254186 at a 1/50 dilution followed by ab150080 at a 1/1000 dilution.

Negative control 2 : ab179513 at a 1/200 dilution followed by ab150113 at a 1/1000 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 19 antibody [LP2K] (AB254186)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Cytokeratin 19 with ab254186 at 1/10000 (0.105 μg/ml) dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human kidney is observed. The section was incubated with ab254186 for 30 mins at room temperature and followed by specific mouse IgG antibody (ab190482) for 8mins. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 19 antibody [LP2K] (AB254186)

Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling Cytokeratin 19 with ab254186 at 1/10000 (0.105 μg/ml) dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The section was incubated with ab254186 for 30 mins at room temperature and followed by specific mouse IgG antibody (ab190482) for 8mins. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Negative control : no staining on human spleen.

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• Flow Cyt

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Flow Cytometry - Anti-Cytokeratin 19 antibody [LP2K] (AB254186)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MCF7 (Human breast adenocarcinoma epithelial cell) cells labelling Cytokeratin 19 with ab254186 at 1/1000 dilution (0.1μg) (Red) compared with a Mouse monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat anti mouse IgG (Alexa Fluor^® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.

• WB

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Western blot - Anti-Cytokeratin 19 antibody [LP2K] (AB254186)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lanes 1-3 : 3 secs; Lane 4 : 48 secs.

All lanes:

Western blot - Anti-Cytokeratin 19 antibody [LP2K] (ab254186) at 1/2000 dilution

Lane 1:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

Human skin tissue lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 44 kDa

Observed band size: 40 kDa

false

• WB

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Western blot - Anti-Cytokeratin 19 antibody [LP2K] (AB254186)

The expression profile observed is consistent with what has been described in the literature (PMID : 24309898).
Negative control : human spleen (PMID : 24309898).

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Cytokeratin 19 antibody [LP2K] (ab254186) at 1/2000 dilution

Lane 1:

Human kidney tissue lysate at 20 µg

Lane 2:

Human colon tissue lysate at 20 µg

Lane 3:

Human stomach tissue lysate at 20 µg

Lane 4:

Human spleen tissue lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 44 kDa

Observed band size: 40 kDa

false