PE Anti-Prostaglandin E Receptor EP2/PTGER2 antibody (ab92755)

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The peptide sequence used to raise the anti-Prostaglandin E Receptor EP4 antibody (ab92763) is SLRTQDATQTSCSTQSDASKQADL and the sequence for the anti-Prostaglandin E Receptor EP2 antibody (ab92755) is GSGRAGPAPKGSSLQVTFPSETLNLSEKCI. These peptides were taken from the residues 459-488 and 335-358 of the human proteins respectively. As you have noted, these regions are both within the cytoplasmic region of the proteins. However, this does not necessarily mean that they would not be suitable for use in Flow Cytometry. It just means that permiabilisation of the cells is likely to be required in order to allow the antibody to reach its target. With these antibodies, we employed 0.1-0.3% Triton X100 to permeabilize the membranes of cells (0.1%) or platelets (0.3%) in order to allow the antibody to penetrate the cell.

I hope this information has been of help. If you require any further information, please do not hesitate to contact us again.

To stain cells, you will need to fix and permeabilize them, since the binding sites are on the cytoplasmic region of the receptor. The only application that has been tested is flow cytometry, and the laboratory that did the validation provide only a very basic protocol. I recommend fixing cells with 4% paraformaldehyde for 20 minutes, rinsing, then permeabilizing them with a solution of detergent, 0.1% - 0.3% Triton X-100 in PBS or TBS. The cells should remain in the detergent solution for at least 10 minutes. Follow this with a blocking step to minimize non-specific interactions of the antibody with other proteins on an in the cells. A 5% solution of normal rabbit serum or BSA in buffer should be sufficient, for 20 minutes. Incubate with the antibody for at least 30 minutes at a concentration of 4 ug/ml. The antibody diluent should include 0.1% Triton X-100, and 5% normal serum or BSA to continue blocking non-specific binding. After washing, the cells are ready for the flow cytometer or observation. If you intend to observe the cells under a fluorescent microscope, please be aware that this application has not been tested. Phycoerythrin is susceptible to photobleaching. Be sure to keep the slides in the dark as much as possible. I hope this helps. If you have any questions, please contact us.

Thank you for contacting us. The immunogen sequence, corresponding to C terminal amino acids 335-358 of Human Prostaglandin E Receptor EP2, is located in the cytoplasm. A map of the protein can be found at the UniProt entry at the following link: http://www.uniprot.org/uniprot/P43116 A BLAST search of the immunogen returns only Prostaglandin E2 receptor EP2 subtype. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.