PTGER3 Antibody / Prostaglandin E2 receptor EP3 (FY13382)

Flow Cytometry analysis of human THP-1 cells using anti-PTGER3 antibody. Overlay histogram showing THP-1 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-PTGER3 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.

Western blot analysis of PTGER3 using anti-PTGER3 antibody. Lane 1: human HEL whole cell lysates, Lane 2: human K562 whole cell lysates, Lane 3: human SH-SY5Y whole cell lysates, Lane 4: human THP-1 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-PTGER3 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. PTGER3 antibody detects a major band at approximately 43 kDa, the expected size of the EP3 receptor. HEL and K562 lysates show a slightly higher migrating band, consistent with documented PTGER3 isoform diversity and cell-type-dependent glycosylation reported for this GPCR.