![All lanes : Anti-Lysosomal acid lipase antibody [9G7F12] (ab36597)Lane 1 : Molecular weight markerLane 2 : Truncated LAL recombinant protein](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_3/18181_ab36597_1.gif)
All lanes : Anti-Lysosomal acid lipase antibody [9G7F12] (ab36597)Lane 1 : Molecular weight markerLane 2 : Truncated LAL recombinant protein
ab36597 (2 µg/ml) staining lysosomal acid lipase in human lung using an automated system (DAKO Autostainer Plus). Using this protocol there is lysosomal staining in the bronchiolar epithelium and resident macrophages .Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH 6.0 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.
![Overlay histogram showing HepG2 cells stained with ab36597 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab36597, 0.5µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_3/18183_Lysosomal-acid-lipase-Primary-antibodies-ab36597-2.jpg)
Overlay histogram showing HepG2 cells stained with ab36597 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab36597, 0.5µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.