Anti-Cytokeratin 18 抗体 [E431-1]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Cytokeratin 18 antibody [E431-1] (AB32118)

Flow cytometry overlay histogram showing left MCF7 positive cells and right negative A375 stained with ab32118 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab32118) (1x 106 in 100μl at 0.008μg/ml (1/258750)) for 30min at 22°C.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C

Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

This antibody gave a positive signal in MCF7 Fixed with 4% formaldehyde (10 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [E431-1] (AB32118)

ab32118 showing positive staining in Normal kidney tissue.

• ICC/IF

PubMed

Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 18 antibody [E431-1] (AB32118)

Indirect immunofluorescence staining with antibodies against DNA, cytokeratin-7 and cytokeratin-18 (upper panel) or against DNA, vimentin and cytokeratin-18 (lower panel). Representatives are presented. Scale bar : 20 μm.

Control or treated cells were fixed for 15 min with 4% PFA containing 0.1% Triton X-100 at room temperature. The following primary antibodies were used for staining : monoclonal mouse antibodies against vimentin and cytokeratin-7 (both 1 : 100, DAKO) and monoclonal rabbit antibody against cytokeratin-18 (1 : 50, Abcam). DNA was stained using DAPI (4',6-diamidino-2-phenylindole-dihydrochlorid) (Roche). Slides were examined using an Axio Imager 7.1 microscope (Zeiss) and images were taken using an Axio Cam MRm camera (Zeiss). The immunofluorescence stained slides were also examined by a confocal laser scanning microscope (CLSM) (Leica CTR 6500, Heidelberg). Images were processed using Photoshop.

Muschol-Steinmetz C. et al PLoS One. 2013 Sep 19;8(9):e73337. doi: 10.1371/journal.pone.0073337. eCollection 2013. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [E431-1] (AB32118)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human stomach tissue sections labeling Cytokeratin 18 with purified ab32118 at 1/500 dilution (0.08 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [E431-1] (AB32118)

ab32118 showing positive staining in Normal colon tissue.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Cytokeratin 18 antibody [E431-1] (AB32118)

Overlay histogram showing MCF7 cells stained with ab32118 (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 (ab32118, 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. Acquisition of >5,000 events was performed. This antibody gave a positive signal in MCF7 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 18 antibody [E431-1] (AB32118)

Immunocytochemistry/Immunofluorescence analysis of HT-29 cells labelling Cytokeratin 18 with purified ab32118 at 1/500. Cells were fixed with 100% Methanol. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (ab150077) at 1/1000 dilution was used as the secondary antibody. The cells were co-stained with ab7291, a mouse anti-tubulin antibody (1/1000) using ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) as the secondary antibody. Nuclei were counterstained with DAPI (blue).

For negative control 1, rabbit primary antibody was used followed by anti-mouse secondary antibody (ab150120).
For negative control 2, mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077) were used.

Alexa Fluor^® 488 (ab194124) conjugated version is available for this clone.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 18 antibody [E431-1] (AB32118)

Immunocytochemistry/Immunofluorescence analysis of HeLa (+ve) and U87-MG (-ve) cells labelling Cytokeratin 18 with ab32118 at 2 ug/ml overnight at +4°C. 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. A preadsorbed Alexa Fluor® 488-conjugated goat anti-rabbit IgG (ab150081) at 1/1000 dilution was used as the secondary antibody. The cells were co-stained with ab7291, a mouse anti-tubulin antibody (1/1000), using ab150119, a preadsorbed Alexa Fluor® 647-conjugated goat anti-mouse IgG (1/1000) as the secondary antibody. Nuclei were counterstained with DAPI (blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 4% formaldehyde (10 min).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [E431-1] (AB32118)

ab32118 showing positive staining in Breast carcinoma tissue.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Cytokeratin 18 antibody [E431-1] (AB32118)

Intracellular Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Cytokeratin 18 with purified ab32118 at 1/20 dilution (2μg/ml) (red). Cells were fixed with 80% Methanol and permeabilised with 0.1% Tween-20. A Goat anti rabbit IgG (Alexa Fluorr^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

R-PE (ab210410) conjugated version is available for this clone.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [E431-1] (AB32118)

Fluorescent immunohistochemical analysis of paraffin-embedded human normal kidney tissue using ab32118. Green-CK18 red-PI

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [E431-1] (AB32118)

ab32118 showing positive staining in Gastric adenocarcinoma tissue.

• WB

Lab

Western blot - Anti-Cytokeratin 18 antibody [E431-1] (AB32118)

All lanes:

Western blot - Anti-Cytokeratin 18 antibody [E431-1] (ab32118) at 1/2000 dilution

Lane 1:

A431 (Human epidermoid carcinoma epithelial cell) whole cell lysates at 15 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 48 kDa

Observed band size: 48 kDa

false

• WB

Unknown

Western blot - Anti-Cytokeratin 18 antibody [E431-1] (AB32118)

All lanes:

Western blot - Anti-Cytokeratin 18 antibody [E431-1] (ab32118) at 1/10000 dilution

All lanes:

A431 cell lysate

Predicted band size: 48 kDa

Observed band size: 48 kDa

false

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [E431-1] (AB32118)

ab32118 showing negative staining in Glioma tissue.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [E431-1] (AB32118)

ab32118 showing negative staining in Normal brain tissue.