Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker

• IHC-Fr

AbReview59038****

Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (AB181598)

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.

Image courtesy of Dr. Isabelle Miletich

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (AB181598)

Intracellular 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (AB181598)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (AB181598)

Immunohistochemical analysis of formalin fixed paraffin embedded human breast labelling cytokeratin 7 with ab181598 at a concentration of 0.1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a ChromoMap DAB kit and anti-rabbit HQ and anti-HQ HRP detection. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins. ab181598 anti-Cytokeratin 7 antibody [EPR17078] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (AB181598)

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.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (AB181598)

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (AB181598)

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (AB181598)

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.

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (AB181598)

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)

Fransen-Pettersson, N. et al PLoS One. 2016 Jul 21;11(7):e0159850. doi: 10.1371/journal.pone.0159850. eCollection 2016 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

• WB

Lab

Western blot - Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (AB181598)

Western blot : Anti-KRT7 antibody [EPR17078] (ab181598) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab181598 was shown to bind specifically to KRT7. A band was observed at 40-55 kDa in wild-type A549 cell lysates with no signal observed at this size in KRT7 knockout cell line. To generate this image, wild-type and KRT7 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (ab181598) at 1/10000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

KRT7 knockout A549 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

MCF7 cell lysate at 20 µg

Observed band size: 40-55 kDa

false

• WB

Supplier Data

Western blot - Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (AB181598)

Blocking/Dilution buffer : 5% NFDM/TBST.

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

All lanes:

Western blot - 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: 42 kDa,45 kDa,51 kDa

false

• WB

Supplier Data

Western blot - Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (AB181598)

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

All lanes:

Western blot - Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (ab181598) at 1/1000 dilution

All lanes:

Mouse liver lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 51 kDa

false

• WB

Supplier Data

Western blot - Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (AB181598)

Blocking/Dilution buffer : 5% NFDM/TBST.

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

All lanes:

Western blot - Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (ab181598) at 1/50000 dilution

Lane 1:

Rat bladder tissue at 10 µg

Lane 2:

Mouse lung tissue 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: 42 kDa,52 kDa

false

• WB

Supplier Data

Western blot - Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (AB181598)

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

All lanes:

Western blot - Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (ab181598) at 1/5000 dilution

All lanes:

Mouse kidney 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: 42 kDa,45 kDa,51 kDa

false