All lanes : Anti-Endothelin A Receptor antibody (ab76259) at 1/500 dilutionLane 1 : Extracts from COS-7 cellsLane 2 : Extracts from COS-7 cells plus 5µg immunizing peptideLysates/proteins at 5 µg per lane.
Immunofluorescence analysis of LOVO cells using ab76259 at a 1/500 dilution.Left image un-treated.Right image treated with immunizing peptide.
ab76259 (4µg/ml) staining Endothelin A receptor in human cerebellum using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of the endothelium.Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. 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, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.