Anti-hair cortex Cytokeratin antibody [AE13] (ab16113)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - hair cortex Cytokeratin antibody [AE13] (ab16113)

ab16113 at 1/100 staining mouse skin tissue sections by IHC-P. The tissue was paraformaldehyde fixed and blocked with BSA. A heat mediated antogen retrieval step was performed. The tissue was incubated with the antibody for 16 hours and then an Alexa-Fluor 488 conjugated goat anti-mouse antibody was used as the secondary. The image shows hair cortex cytokeratin staining in green with DAPI nuclei counterstain in blue.

This image is courtesy of an anonymous Abreview

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - hair cortex Cytokeratin antibody [AE13] (ab16113)

ab16113 staining hair cortex Cytokeratin in adult mouse nail by Immunohistochemistry (paraffin embedded sections). Tissue was fixed with paraformaldehyde and blocked with 10% serum for 1 hour at 20°C followed by incubation with the primary antibody, at a 1/200 dilution, for 12 hours at 4°C. A Biotin-conjugated goat anti-mouse monoclonal was used as secondary antibody at a 1/400 dilution.

This image is courtesy of an anonymous abreview.

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  Western blot - hair cortex Cytokeratin antibody [AE13] (ab16113)

All lanes : Anti-hair cortex Cytokeratin antibody [AE13] (ab16113) at 1 µg/ml

Lane 1 : Skin (Mouse) Whole Cell Lysate - normal tissue, 14 days old (ab7271)
Lane 2 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate

Lysates/proteins at 10 µg per lane.

Secondary
Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed (ab97040) at 1/5000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 44 - 46 kDa
Observed band size : 44 kDa (why is the actual band size different from the predicted?)
Additional bands at : 25 kDa,55 kDa. We are unsure as to the identity of these extra bands.

Exposure time : 20 minutes

  Flow Cytometry - hair cortex Cytokeratin antibody [AE13] (ab16113)

Overlay histogram showing A431 cells stained with ab16113 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab16113, 1/20 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells ) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in A431 cells fixed with 100% methanol (5 min)/permeabilized in 0.1% PBS-Triton X-100 used under the same conditions.