PNLIP Antibody / Pancreatic Lipase (FY12082)

Immunofluorescent staining of PNLIP using anti-PNLIP antibody (green). PNLIP was detected in an immunocytochemical section of U2OS cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 ug/ml rabbit anti-PNLIP antibody overnight at 4oC. DyLight 488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37oC. The section was counterstained with DAPI (blue). Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of Pancreatic Lipase using anti-PNLIP antibody. Lane 1: rat pancreas tissue lysates, Lane 2: mouse pancreas tissue lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-PNLIP antibody at 0.5 ug/ml overnight at 4oC, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal was developed using enhanced chemiluminescent. The expected band size for PNLIP is at 51 kDa and the protein is often observed as several closely migrating glycoforms. This is more pronounced in rodent samples than human samples.

Flow Cytometry analysis of U251 cells using anti-PNLIP antibody. Overlay histogram showing U251 cells stained with (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-PNLIP antibody (1 ug/million cells) for 30 min at 20oC. DyLight 488 conjugated goat anti-rabbit IgG (5-10 ug/million cells) was used as secondary antibody for 30 minutes at 20oC. Isotype control antibody (Green line) was rabbit IgG (1 ug/million cells) used under the same conditions. Unlabelled sample (Red line) was also used as a control.