Cervix consists of two distinct KRT5+ stratified and KRT7+/8+ columnar epithelial lineages.

Panel A shown only.

Transition zone (TZ) including stratified and columnar epithelium from human (A,C) and mouse (B, D) cervix tissue sections immunolabeled with antibodies against KRT5 (ab193894) and KRT7 (ab209601); nuclei are shown in blue.

Organoids were washed five times with cold PBS to remove Matrigel before fixing with 4% paraformaldehyde for 1 h at room temperature (RT) followed by washing with PBS twice. Organoids were then subjected to dehydration in an ascending ethanol series followed by isopropanol and acetone for 20 min each. The dehydrated organoids were paraffin-embedded and 5 µM sections cut on a Microm HM 315 microtome. Mouse and human tissues were extensively washed with PBS and fixed using 4% PFA overnight at RT. Samples were subjected to dehydration in an ascending ethanol series followed by isopropanol and xylene (60 min each) followed by paraffinization using a Leica TP1020 tissue processor. The tissue was embedded and 5 µM sections cut on a microtome. For immunostaining, paraffin sections were deparaffinized and rehydrated, followed by treatment with antigen retrieval solution. Sections were blocked using blocking buffer (1% BSA and 2% FCS in PBS) for 1 h at RT. Primary antibodies were diluted in blocking buffer and incubated for 90 mins at RT followed by five PSB washes before 1 h incubation with secondary antibodies diluted in blocking buffer along with Hoechst or Draq5. Sections were washed with PBS five times and mounted. Images were acquired with a Leica TCS SP8 confocal microscope.

ab209601 staining Cytokeratin 7 in A549 cells. The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab209601 at 1/100 dilution (pseudocolored in green) and ab195884, Rat monoclonal to Tubulin (Alexa Fluor^® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in A549 cells fixed with 4% formaldehyde (10 min).