All lanes : Anti-NEK9 (phospho T210) antibody (ab63553) at 1/500 dilutionLane 1 : extracts from HepG2 cells at 30 µg/mlLane 2 : extracts from HepG2 cells at 30 µg with immunizing peptide at 10 µg
ab63553 at 1/500 dilution staining HeLa cells, without (left) and with (right) immunizing peptide.
Ab63553 staining Human colon. Staining is localised to the cytoplasm and nucleus.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 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.