Anti-Cytokeratin 18 antibody [SP69]

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Cytokeratin 18 antibody [SP69] (AB93741)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast labelling Cytokeratin 18 with ab93741 at 1/200 dilution (1.02 µg/mL) (B), Collagen VI with ab271938 at 1/500 dilution (2.084 μg/ml) (C) and Cytokeratin 14 with ab236439 at 1/2000 dilution ( 0.519 μg/ml) (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. Panel A : merged staining of anti-Cytokeratin 14 (magenta; Opal™690), anti-Cytokeratin 18 (green; Opal™520) and anti-Collagen VI (red; Opal™570) on human breast. Panel B : anti-Cytokeratin 18 stained on luminal epithelial cells. Panel C : anti-Collagen VI stained on stroma. Panel D : anti-Cytokeratin 14 stained on myoepithelial cells. The section was incubated in three rounds of staining : in the order of ab236439 for 30 mins, ab93741 for 10 mins, and ab271938 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Cytokeratin 18 antibody [SP69] (AB93741)

Flow cytometric analysis of rabbit anti-Cytokeratin 18 (SP69) antibody ab93741(1/100) in HeLa cells (green) compared to negative control of rabbit IgG (blue).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Cytokeratin 18 antibody [SP69] (AB93741)

Flow Cytometry analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling Cytokeratin 18 with purified ab93741 at 1 : 240 dilution (1.00 µg/ml) Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1 : 2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / Blue.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [SP69] (AB93741)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human endometrium tissue sections labeling Cytokeratin 18 with ab93741 at 1/200 dilution (1.21 µg/ml). Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 10 mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [SP69] (AB93741)

Immunihistochemical staining of human stomach with ab93741.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 18 antibody [SP69] (AB93741)

ICC/IF image of ab93741 stained HCT116 cells. The cells were 4% formaldehyde fixed (10 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 ab93741 at 1/500 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 Goat anti-Rabbit IgG (H+L) used at a 1/250 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/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 18 antibody [SP69] (AB93741)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling Cytokeratin 18 with purified ab93741 at 1 : 100(2.4 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1 : 200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [SP69] (AB93741)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human stomach tissue sections labeling Cytokeratin 18 with ab93741 at 1/200 dilution (1.21 µg/ml). Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 10 mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Cytokeratin 18 antibody [SP69] (AB93741)

Overlay histogram showing MCF7 cells stained with ab93741 (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 (ab93741, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. 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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [SP69] (AB93741)

ab93741 at 1/200 dilution staining Cytokeratin 18 in formalin-fixed, paraffin-embedded Human breast carcinoma tissue.

• WB

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Western blot - Anti-Cytokeratin 18 antibody [SP69] (AB93741)

All lanes:

Western blot - Anti-Cytokeratin 18 antibody [SP69] (ab93741) at 1/25 dilution

All lanes:

A431 cell lysate

Predicted band size: 48 kDa

Observed band size: 52 kDa

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