Ab48614 staining human tonsil. Staining is localized to the cytoplasm.Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.Sections were stained using an automated system (Dako PT Link), at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer, EDTA pH 9.0. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
Anti-Ceruloplasmin antibody (ab48614) at 1 µg/ml + Human Plasma Total Protein Lysate at 10 µgSecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
IHC-FoFr image of ceruloplasmin staining on mouse duodenal sections using ab48614. These sections were incubated in triton X (0.3% in 0.1% PBS) to permeabilise the cells and in 10% serum for 1h to block non-specific protein-protein interactions. The sections were then incubated with the antibody ab48614(1:200) overnight at +4°C. Specific protien binding was visualised using an Alexa 488 conjugated donkey secondary antibody (green staining). Alexa 568 conjugated secondary was used to visualise ab55027 binding to ferritin (red staining). See Abreview