IHC image of pSer26 Amyloid beta staining in a section of frozen Human Alzheimer brain*.

The section was fixed using 10% formaldehyde in 1XPBS for 10 minutes. No antigen retrieval step was performed prior to staining. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M 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 ab219818 at 1µg/ml and ab2539 (Rabbit polyclonal Antibody to beta Amyloid) at 1/100. The section was then incubated with ab150119 (Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^®647, 1/1000)) (shown in red) and ab150088 (Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) preadsorbed, 1/1000) (shown in green) for 1 hour at room temperature. The secondary-only control insert image is taken from an identical assay without primary antibody. The section was then mounted using Fluoromount^®.

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

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

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

Phospho- or control peptides (1 microgram per mL) were immobilised to a plate and blocked with 5% BSA (PBS-T) before incubation with ab219818 for 1 hour at room temperature. Antibody binding was detected by Goat anti-Mouse IgG (HRP) secondary antibody and signal developed by TMB substrate.