Anti-Cytokeratin 15 antibody [LHK15]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 15 antibody [LHK15] (AB80522)

IHC image of Cytokeratin 15 staining in a formalin fixed, paraffin embedded normal human breast tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab80522, 5μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

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

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 15 antibody [LHK15] (AB80522)

ab80522 staining Cytokeratin 15 in A431 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-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 ab80522 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 100% methanol (5 min).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• ICC/IF

AbReview15530****

Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 15 antibody [LHK15] (AB80522)

ab8969 staining keratin 15 in rat hair folicles by immunocytochemistry/ immunofluorescence. Cells were paraformaldehyde fixed and permeabilized in 0.25% Triton X-100 prior to blocking in 1% BSA for 10 minutes at 21°C. The primary antibody was diluted 1/100 and incubated with the sample for 1 hour at 21°C. A TRITC conjugated mouse anti-mouse antibody, diluted 1/85, was used as the secondary.

This image is courtesy of an Abreview submitted by Maciej Gagat

• IHC-P

AbReview40266****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 15 antibody [LHK15] (AB80522)

ab80522 staining Cytokeratin 15 in mouse colon tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde; antigen retrieval was by heat mediation. Samples were incubated with primary antibody (undiluted) for 10 hours.

This image is courtesy of an anonymous Abreview