All lanes : Anti-C5R1 antibody (ab59390) at 1/500 dilutionLane 1 : Extracts from HeLa cells, treated with PMA (125ng/ml, 30mins).Lane 2 : Extracts from HeLa cells, treated with PMA (125ng/ml, 30mins) plus peptide.
ab59390 staining C5R1 in human lung.Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.Sections were stained using an automated system (DAKO Autostainer Plus), at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers EDTA pH 9.0. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min 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.