Anti-Cytokeratin 7 antibody [EPR17078] - BSA and Azide free

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Cytokeratin 7 antibody [EPR17078] - BSA and Azide free (AB220804)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181598).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 7 antibody [EPR17078] - BSA and Azide free (AB220804)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181598).

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

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 7 antibody [EPR17078] - BSA and Azide free (AB220804)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181598).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 7 antibody [EPR17078] - BSA and Azide free (AB220804)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181598).

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] - BSA and Azide free (AB220804)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181598).

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] - BSA and Azide free (AB220804)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181598).

• WB

Lab

Western blot - Anti-Cytokeratin 7 antibody [EPR17078] - BSA and Azide free (AB220804)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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 (<a href='/en-us/products/primary-antibodies/cytokeratin-7-antibody-epr17078-cytoskeleton-marker-ab181598'>ab181598</a>) 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

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