HRP Anti-Cytokeratin 7 antibody [EPR17078]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-Cytokeratin 7 antibody [EPR17078] (AB209945)

IHC image of Cytokeratin 7 staining in a section of formalin-fixed paraffin-embedded normal human breast*, performed on a Leica BOND™. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab209945, 1/100 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• WB

Lab

Western blot - HRP Anti-Cytokeratin 7 antibody [EPR17078] (AB209945)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab209945 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

All lanes:

Western blot - HRP Anti-Cytokeratin 7 antibody [EPR17078] (ab209945) at 1/5000 dilution

Lane 1:

Lung (Mouse) Tissue Lysate at 10 µg

Lane 2:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Predicted band size: 51 kDa

Observed band size: 53 kDa

true

Exposure time: 15s

• WB

Lab

Western blot - HRP Anti-Cytokeratin 7 antibody [EPR17078] (AB209945)

ab209945 was shown to specifically react with KRT7 in wild-type A549 cells as signal was lost in KRT7 knockout cell line ab261867 (knockout cell lysate ab261676). Wild-type and KRT7 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. ab209945 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibody at 1/20000 dilution for 1 hour at room temperature. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.

All lanes:

Western blot - HRP Anti-Cytokeratin 7 antibody [EPR17078] (ab209945) at 1/5000 dilution

Lane 1:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human KRT7 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-krt7-knockout-a549-cell-line-ab261867'>ab261867</a>) at 20 µg

Lane 3:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 4:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Predicted band size: 51 kDa

false