IHC image of ab12077 staining in 100% methanol fixed frozen tissue section of human skin.

Non-specific protein-protein interactions were blocked using TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 3% (w/v) BSA for 1 hour at room temperature. The section was then incubated with ab12077 (1μg/ml dilution) in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA overnight at +4°C. The section was then incubated with ab150119 (Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 647)) and DAPI for 1 hour at room temperature.

The DAPI only control (no antibody) inset shows no autofluorescence, demonstrating that any Alexa Fluor^® 647 signal is derived directly from bound ab12077. 

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

ab12077 staining Desmoglein 1/DSG1 in human skin tissue by Immunohistochemistry (Frozen sections).Tissue was fixed in methanol, permeabilized using 0.4% Triton X-100, blocked with 10% serum for 30 minutes at 25°C, then incubated with ab12077 at 5µg/ml for 16 hours at 4°C. The secondary used was an Alexa-Fluor 594 conjugated goat anti-mouse polyclonal (highly adsorbed), used at a 1/1000 dilution.

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