All lanes : Anti-GAB1 antibody (ab47513)Lane 1 : HUVEC cell extract treated with EGF (200ng/ml, 10 min)Lane 2 : HUVEC cell extract treated with EGF (200ng/ml, 10 min) with peptideLysates/proteins at 30 µg per lane.
Ab47513 staining human normal tonsil. Staining is localised to the cytoplasm and cell membrane.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 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.