IHC image of Podoplanin / gp36 staining in a section of formalin-fixed paraffin-embedded normal mouse skin. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6). Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 1% (w/v) BSA for 1h at room temperature. The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab11936 at 1/250. The secondary antibody (shown in green) was ab180063 at 1/500 (Goat Anti-Syrian hamster IgG H&L (Alexa Fluor® 488) and counterstained using ab190573, Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker (Alexa Fluor® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Fluoromount^®.

The negative control (inset) is a secondary-only assay to demonstrate low non-specific binding of the secondary antibody.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

ab11936 staining Podoplanin / gp36 in murine skin tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
Tissue was fixed in paraformaldehyde and an enzymatic antigen retrieval step was performed using Protein K. Samples were then blocked with 5% serum for 30 minutes at room temperature and then incubated with ab11936 at a 1/400 dilution for 16 hours at 4°C. The secondary used was a biotin conjugated goat anti-hamster IgG used at a 1/500 dilution.

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