All lanes : Anti-Casein Kinase 1 alpha antibody (ab63373) at 1/500 dilutionLane 1 : HUVEC cell extractLane 2 : K562 cell extractLane 3 : Peptide blocking experiment, K562 LysateLysates/proteins at 30 µg per lane.
Immunofluorescence analysis of HeLa cells, using ab63373 at 1/500 dilution. Right hand panel was treated with immunising peptide.
Ab63373 staining human normal lung. Staining is localised to the cytoplasm (apical).Left panel: with primary antibody at 2 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 antigen retrieval buffer EDTA pH 9.0 in a DAKO PT Link. 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.