ab28484 (4µg/ml) staining Lubricin/MSF in Human liver using an automated system. Using this protocol there is staining of the cytoplasm and nuclei in hepatic arteries and plates of hepatic cells.
Sections were rehydrated and antigen retrieved with  buffers EDTA pH 9.0 . Slides were peroxidase blocked in 3% H₂O₂ in methanol for 10 mins. They were then blocked  for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected  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, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

Primary antibody was diluted in antibody diluent using a Fast Western Blot kit and followed by the optimized HRP reagent. Chemiluminescent detection was performed using SuperSignal West Dura.