S100P Antibody for IF / S100 Calcium Binding Protein P Antibody for Immunofluorescence (FY12704)

S100P Antibody for IF in cultured cells. Immunofluorescence analysis using S100P Antibody for IF (green) and beta tubulin antibody (red) demonstrates S100 Calcium Binding Protein P / S100P staining predominantly within nuclear regions of the cells, showing punctate signal within the nuclei. Beta tubulin staining outlines the cytoplasmic cytoskeleton and defines overall cell morphology. Cells were incubated with rabbit anti-S100P antibody and mouse anti-beta tubulin antibody followed by DyLight 488 goat anti-rabbit IgG and DyLight 594 goat anti-mouse IgG secondary antibodies. Fluorescent imaging was performed using a fluorescence microscope with appropriate filter sets for the fluorophores.

Western blot analysis of S100P using anti-S100P antibody. Lane 1: human RT4 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-S100P 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. A predominant band is detected at ~14 kDa, which is typical for S100P due to anomalous SDS-PAGE mobility of EF-hand S100 proteins relative to the ~10 kDa calculated mass.

Flow Cytometry analysis of SiHa cells using anti-S100P antibody. Overlay histogram showing SiHa 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-S100P 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.