Overview

  • Product name
    Anti-Prostaglandin E Receptor EP4 antibody
    See all Prostaglandin E Receptor EP4 primary antibodies
  • Description
    Rabbit polyclonal to Prostaglandin E Receptor EP4
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Rat, Chimpanzee
  • Immunogen

    Synthetic peptide corresponding to Human Prostaglandin E Receptor EP4 aa 1-100 conjugated to keyhole limpet haemocyanin.
    Database link: P35408
    (Peptide available as ab45294)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: Raji and Ramos

Properties

Applications

Our Abpromise guarantee covers the use of ab45295 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 73 kDa (predicted molecular weight: 53 kDa).

Target

  • Function
    Receptor for prostaglandin E2 (PGE2). The activity of this receptor is mediated by G(s) proteins that stimulate adenylate cyclase. Has a relaxing effect on smooth muscle. May play an important role in regulating renal hemodynamics, intestinal epithelial transport, adrenal aldosterone secretion, and uterine function.
  • Tissue specificity
    High in intestine and in peripheral blood mononuclear cells; low in lung, kidney, thymus, uterus, vasculature and brain. Not found in liver, heart, retina oe skeletal muscle.
  • Sequence similarities
    Belongs to the G-protein coupled receptor 1 family.
  • Cellular localization
    Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • EP 4 antibody
    • EP4 antibody
    • EP4R antibody
    • MGC126583 antibody
    • PE2R4_HUMAN antibody
    • PGE receptor EP4 subtype antibody
    • PGE2 receptor EP4 subtype antibody
    • Prostaglandin E receptor 4 antibody
    • Prostaglandin E receptor 4 subtype EP4 antibody
    • Prostaglandin E2 receptor antibody
    • Prostaglandin E2 receptor EP4 subtype antibody
    • Prostanoid EP4 receptor antibody
    • PTGER 2 antibody
    • PTGER 4 antibody
    • PTGER2 antibody
    • PTGER4 antibody
    see all

Images

  • ICC/IF image of ab45295 stained HeLa 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 (ab45295, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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. This antibody also gave a positive result in 4% PFA fixed (5 min) HeLa cells at 5µg/ml.
  • Anti-Prostaglandin E Receptor EP4 antibody (ab45295) at 1 µg/ml + Ramos (Human Burkitt's lymphoma cell line) Whole Cell Lysate at 20 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size: 53 kDa
    Observed band size: 73 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 30 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 20 minutes


    ab45295 detects a 73 kDa band in Western Blot which we believe corresponds to the glycosylated form of the protein.

References

This product has been referenced in:
  • Wang J  et al. Activation of PGE2/EP2 and PGE2/EP4 signaling pathways positively regulate the level of PD-1 in infiltrating CD8+T cells in patients with lung cancer. Oncol Lett 15:552-558 (2018). Read more (PubMed: 29285200) »
See 1 Publication for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Application
Western blot
Sample
Human Tissue lysate - whole (Abdominal endometriotic lesion)
Gel Running Conditions
Reduced Denaturing (12% gel)
Loading amount
15 µg
Specification
Abdominal endometriotic lesion
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Mar 22 2017

Answer

Thank you for your reply.

You would probably be best suited using a kit that is able to isolate the membrane fraction completely if you are looking at a membrane protein like Prostaglandin E Receptor EP4. You may want to use a kit like our ab139409, membrane fractionation kit. https://www.abcam.com/membrane-fractionation-kit-ab139409.html

I hope this information helps. Please contact us with any other questions.

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Answer

Thank you for this feedback! I am happy to hear that it now works :-)

By the way, if you like you could submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates. To find out more about our Abreview system, please see the following link: https://www.abcam.com/abreviews

Have a great weekend!

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Answer

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this vial of antibody.

The details you have kindly provided will provide us with vital information for our monitoring of product quality.

I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Reviewing the details, I can suggest you have regrettably received a bad vial.

I apologise for the inconvenience and am pleased to offer you a free of charge replacement (we only have lot 949491 in stock) or credit note in compensation.

Could you please confirmthat theoriginal order number was xxxx (21.January 2012)?

In addition, I can suggest you may like to consider checking if the secondary antibody is still working.

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

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Question
Answer

For a Jurkat cell lysate for western blotting, I recommend item ab7899:

Click here (or use the following: https://www.abcam.com/index.html?datasheet=7899).

The lysate is presented in SDS-PAGE loading buffer:

50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue, containing 5% b-mercaptoethanol.

If you have any questions, please contact me.

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Answer

Thank you for contacting us. Your customer can use a Jurkat cell lysate as a positive control. We have the lysate as ab7899: Click here (or use the following: https://www.abcam.com/index.html?datasheet=7899). The Jurkat cell lysate is also an appropriate positive control for the other Anti-Prostaglandin E Receptor EP4 antibody you asked about, ab45295. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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