Anti-Lysosomal acid lipase/LAL antibody [9G7F12]

• Flow Cyt

Unknown

Flow Cytometry - Anti-Lysosomal acid lipase/LAL antibody [9G7F12] (AB36597)

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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lysosomal acid lipase/LAL antibody [9G7F12] (AB36597)

ab36597 (2 μg/ml) staining Lysosomal acid lipase/LAL 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.

• WB

Unknown

Western blot - Anti-Lysosomal acid lipase/LAL antibody [9G7F12] (AB36597)

All lanes:

Western blot - Anti-Lysosomal acid lipase/LAL antibody [9G7F12] (ab36597)

All lanes:

Truncated LAL recombinant protein

Predicted band size: 45 kDa

Observed band size: 45 kDa

false