All lanes : Anti-Prostaglandin E Receptor EP2/PTGER2 antibody [EPR8030(B)] (ab167171) at 1/1000 dilution

Lane 1 : Mouse kidney tissue lysate
Lane 2 : Mouse spleen tissue lysate
Lane 3 : Rat spleen tissue lysate
Lane 4 : MDBK (cow kidney) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 40 kDa

Blocking and diluting buffer: 5% NFDM/TBST

ab167171 staining Prostaglandin E Receptor EP2/PTGER2 in the WI-38 (Human Fetal Lung Fibroblast) cell line by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/80. A goat anti rabbit IgG (Alexa Fluor^® 488) at a dilution of 1/2000 was used as the secondary antibody.

Isoytype control: Rabbit monoclonal IgG (Black)

Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

ab167171 staining Prostaglandin E Receptor EP2/PTGER2 in WI-38 (Human Fetal Lung Fibroblast) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/250. A goat anti rabbit IgG (Alexa Fluor^® 488) (ab150077) was used as the secondary antibody at 1/1000. ab7291 and ab150120 were used as counterstains for primary antibody ab167171 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.

Negative control 1: Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
Negative control 2: Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077)

ab167171 staining Prostaglandin E Receptor EP2/PTGER2 in human placenta tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/200. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at 1/500.

Negative control 1: PBS in place of primary antibody.

All lanes : Anti-Prostaglandin E Receptor EP2/PTGER2 antibody [EPR8030(B)] (ab167171) at 1/1000 dilution

Lane 1 : Mouse kidney tissue lysate
Lane 2 : WI-38 (human lung) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 40 kDa

Blocking and diluting buffer: 5% NFDM/TBST

ab167171 stained SV40LT-SMC (rat) cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab167171 at 1/200 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

All lanes : Anti-Prostaglandin E Receptor EP2/PTGER2 antibody [EPR8030(B)] (ab167171) at 1/1000 dilution

Lane 1 : Human placenta lysates
Lane 2 : Human fetal lung lysates
Lane 3 : JAR cell lysates
Lane 4 : WI38 cell lysates

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP labeled goat anti-rabbit at 1/2000 dilution

Predicted band size: 40 kDa

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Prostaglandin E Receptor EP2/PTGER2 with ab167171 at 1/250 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.