Anti-Cytokeratin 19 antibody [A53-B/A2] - Cytoskeleton Marker

• IHC-P

AbReview19313****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 19 antibody [A53-B/A2] - Cytoskeleton Marker (AB7754)

ab7754 staining Cytokeratin 19 in Human liver tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 5% milk for 30 minutes at 37°C; antigen retrieval was by heat mediation in citrate buffer. Samples were incubated with primary antibody (1/1000 in antibody diluent) for 1 hour at 37°C. An undiluted HRP-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

This image is courtesy of an anonymous Abreview

• ICC/IF

PubMed

Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 19 antibody [A53-B/A2] - Cytoskeleton Marker (AB7754)

Immunofluorescence analysis of 1 month hepato-differentiated Human dental pulp stem cells, staining Cytokeratin 19 with ab7754 at 1/60 dilution. A FITC-conjugated anti-mouse IgG was used as the secondary antibody.

Image from Ferro F et al., PLoS One. 2012;7(7):e41774. Epub 2012 Jul 23. Fig 3.; doi:10.1371/journal.pone.0041774; July 23, 2012, PLoS ONE 7(7): e41774.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 19 antibody [A53-B/A2] - Cytoskeleton Marker (AB7754)

ab7754 staining Cytokeratin 19 in human skin.
Left panel : with primary antibody at 2 ug/ml. Right panel : isotype control.
Sections were stained using an automated system (DAKO Autostainer Plus), at room temperature : sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate pH6.1. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 19 antibody [A53-B/A2] - Cytoskeleton Marker (AB7754)

Immunohistochemical analysis of paraffin-embedded human liver tissue stained for Cytokeratin 19 using ab7754 at a 1/100 dilution followed by a GAM IgG-Alexa Fluor^®488 diluted at 1/200 (green). Cell nuclei stained with PI (1 μg/ml; orange).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 19 antibody [A53-B/A2] - Cytoskeleton Marker (AB7754)

ICC/IF image of ab7754 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab7754 at 5μg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.

• ICC

Lab

Immunocytochemistry - Anti-Cytokeratin 19 antibody [A53-B/A2] - Cytoskeleton Marker (AB7754)

ab7754 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 ab7754 at 1/500 dilution 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).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Cytokeratin 19 antibody [A53-B/A2] - Cytoskeleton Marker (AB7754)

Overlay histogram showing MCF7 cells stained with ab7754 (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 (ab7754, 0.5μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Cytokeratin 19 antibody [A53-B/A2] - Cytoskeleton Marker (AB7754)

Separation of MCF-7 cells (red-filled) from human leukocytes (black-dashed) in flow cytometry analysis (intracellular staining) of peripheral whole blood spiked with MCF-7 cells stained using ab7754 (concentration in sample 3 μg/ml, GAM APC).

• WB

Supplier Data

Western blot - Anti-Cytokeratin 19 antibody [A53-B/A2] - Cytoskeleton Marker (AB7754)

Western blotting analysis of human cytokeratin 19 using ab7754 at 2 μg/ml on lysates of HT-29 (Human colorectal adenocarcinoma cell line) cell line and Jurkat (Human T cell leukemia cell line from peripheral blood) cell line (cytokeratin non-expressing cell line; negative control) under non-reducing and reducing conditions.

IRDye800-conjugated anti-mouse IgG1 secondary antibody.

A specific band was detected for cytokeratin 19 at approximately 40 kDa.

All lanes:

Western blot - Anti-Cytokeratin 19 antibody [A53-B/A2] - Cytoskeleton Marker (ab7754)

Predicted band size: 44 kDa

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