PDLIM4 Antibody / PDZ and LIM domain protein 4 (FY12627)

Immunofluorescent staining of PDLIM4 using anti-PDLIM4 antibody (green). PDLIM4 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-PDLIM4 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 nuclear stain (blue). Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of PDLIM4 using anti-PDLIM4 antibody. Lane 1: human Hacat whole cell lysates, Lane 2: human U251 whole cell lysates, Lane 3: human whole cell lysates, Lane 4: human U20S whole cell 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-PDLIM4 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 molecular weight of PDLIM4 is at 35 kDa.

Flow Cytometry analysis of cells using anti-PDLIM4 antibody. Overlay histogram showing cells stained with (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PDLIM4 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Western blot analysis of PDLIM4 using anti-PDLIM4 antibody. Lane 1: human U251 whole cell lysates, Lane 2: human U20S whole cell 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-PDLIM4 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-DyLight 647 Conjugated secondary antibody at a dilution of 1:2000 for 1.5 hour at RT. The expected molecular weight of PDLIM4 is at 35 kDa.

Immunoprecipitating (IP) PDLIM4 in U251 whole cell lysate. Western blot analysis of PDLIM4 using anti-PDLIM4 antibody; Lane 1: U251 whole cell lysates (30ug); Lane 2: Rabbit control IgG instead of anti-PDLIM4 antibody in U251 whole cell lysate; Lane 3: anti-PDLIM4 antibody (2ug) + U251 whole cell lysate (500ug). After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-PDLIM4 antibody at a dilution of 0.5 ug/ml and probed with a goat anti-rabbit IgG-HRP secondary antibody. The signal is developed using ECL Plus Western Blotting Substrate. The expected molecular weight of PDLIM4 is at 35 kDa.