Ab1907 staining Human liver. Staining is localised to cytoplasmic and extracelular regions.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 citrate 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 we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
ICC/IF image of ab1907 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab1907, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
IHC-FoFr image of Apolipoprotein E staining on choroid plexus of APP transgenic mouse brain sections using ab1907 (1:500). The mouse was perfused with 4% PFA in 0.1M PBS. The sections were permeabilised using 0.1% TriktonXin 0.1% PBS. 10% serum was used for 1 hour at 24°C for blocking step. The sections were stained using ab1907 at 1:500 dilution and Rabbit monoclonal to anti-mouse IgG conjugated to Alexa Fluor 568 was used at 1:1000 to detect the primary antibody. DAPI was used to stain the nucleiSee Abreview