All lanes : Anti-Nicotinic Acetylcholine Receptor alpha 4 antibody (ab41172) at 1/800 dilutionLane 1 : SW480 (Human colon adenocarcinoma cell line) Whole Cell Lysate Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lysates/proteins at 10 µg per lane.SecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
ab41172 (4µg/ml) staining Nicotinic Acetylcholine Receptor alpha 4 in human cerebellum using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of nuclear and membrane compartment within the glia region of cellsSections 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.