Hepatic inflammation and abnormal intrahepatic bile ducts in the N-IF mouse

Immunofluorescence staining of bile ducts (CK-7, red), macrophages (F4/80, light blue) and nuclei (DAPI, blue) in livers of 8 week old N-IF and 24αβNOD control mice. Representative images from two independent experiments with a total of six mice are shown. Scale bars are 100 μm.

Cytokeratin is detected with ab181598 at1/1500 dilution in formalin-fixed, paraffin-embedded mouse liver tissue. The secondary antibody is an Anti-Rabbit AlexaFluor^®594 conjugate (red).

(From Figure 1D of Fransen-Pettersson, N. et al)

Immunohistochemical analysis of paraffin-embedded Human mammary gland tissue labeling Cytokeratin 7 with ab181598 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500 dilution. Membrane and cytoplasmic staining on epithelial cells of breast tissues is observed. Counter stained with Hematoxylin.

Negative control: Using PBS instead of primary antibody.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 A549  (Human lung carcinoma) cells  labeling Cytokeratin 7 with ab181598 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Cytoplasmic staining on A549 cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows;
1. ab181598 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

All lanes : Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (ab181598) at 1/50000 dilution

Lane 1 : Rat bladder tissue
Lane 2 : Mouse lung tissue

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 51 kDa
Observed band size: 52, 42 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

 

~45 and 42kDa bands are supported by the literature and competitor’s products, which may be the isoforms.

Immunohistochemical analysis of formaldehyde-fixed, frozen section of mouse salivary gland tissue labeling Cytokeratin 7 with ab181598 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG AlexaFluor^®660.

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Flow cytometry analysis of A549 (human lung carcinoma) cells labeling Cytokeratin 7 (red) with ab181598 at a 1/1500 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with primary and secondary antibodies.

Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (ab181598) at 1/1000 dilution + Mouse liver lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 51 kDa

~45 and 42kDa bands are supported by the literature and competitor’s products, which may be the isoforms.

Immunohistochemical analysis of frozen Mouse kidney tissue labeling Cytokeratin 7 with ab181598 at 1/8000 dilution, followed by Donkey anti-rabbit Alexa Fluor 594 at 1/1000 dilution. Cytoplasmic staining on collecting tube is observed. This data is from our collaborator Dr. Hai Song’s lab (Life Sciences Institute Zhejiang University). Counter stained with DAPI.

Negative control: Using PBS instead of primary antibody.

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Cytokeratin 7 with ab181598 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500 dilution. On mouse liver, only bile duct epithelia are positive, no reaction in hepatocytes is observed. Counter stained with Hematoxylin.

Negative control: Using PBS instead of primary antibody.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

All lanes : Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (ab181598) at 1/20000 dilution

Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) cell lysate at 20 µg
Lane 2 : Rat kidney tissue lysate at 10 µg

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 51 kDa
Observed band size: 51, 45, 42 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

 

~45 and 42kDa bands are supported by the literature and competitor’s products, which may be the isoforms.

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Cytokeratin 7 with ab181598 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500 dilution. On rat liver, only bile duct epithelia are positive, no reaction in hepatocytes is observed. Counter stained with Hematoxylin.

Negative control: Using PBS instead of primary antibody.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (ab181598) at 1/5000 dilution + Mouse kidney lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 51 kDa
Observed band size: 51, 45, 42 kDa why is the actual band size different from the predicted?

~45 and 42kDa bands are supported by the literature and competitor’s products, which may be the isoforms.