Anti-G-protein coupled receptor 30 antibody (ab39742)

Rabbit polyclonal G-protein coupled receptor 30 antibody. Validated in WB, IHC, ICC/IF and tested in Mouse, Rat, Human. Cited in 27 publication(s). Independently reviewed in 7 review(s).

Overview

  • Product name
    Anti-G-protein coupled receptor 30 antibody
    See all G-protein coupled receptor 30 primary antibodies
  • Description
    Rabbit polyclonal to G-protein coupled receptor 30
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-Fr, WB, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 350 to the C-terminus of Human GPCR GPR30.

    Read Abcam's proprietary immunogen policy (Peptide available as ab41565.)

  • Positive control
    • ab39742 gives a positive signal in Human Brain Membrane Lysate, Human Brain Tissue Lysate (data not shown), and MCF7 Whole Cell Lysate (data not shown) This antibody gave a positive signal in the following cell types: U-87 MG.

Properties

Applications

Our Abpromise guarantee covers the use of ab39742 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
IHC-Fr 1/200 - 1/400.
WB 1/250. Detects a band of approximately 55 kDa (predicted molecular weight: 42 kDa).
ICC/IF Use a concentration of 5 µg/ml.
IHC-P Use at an assay dependent concentration. PubMed: 24421881

Target

  • Function
    Receptor for estrogen.
  • Tissue specificity
    Ubiquitously expressed, but is most abundant in placenta. In brain regions, expressed as a 2.8 kb transcript in basal forebrain, frontal cortex, thalamus, hippocampus, caudate and putamen.
  • Sequence similarities
    Belongs to the G-protein coupled receptor 1 family.
  • Cellular localization
    Cell membrane. Endoplasmic reticulum membrane. Golgi apparatus membrane. Protein has been detected in the cell membrane, endoplasmic reticulum and Golgi apparatus. It is currently unclear whether this is a cell surface or intracellular receptor.
  • Information by UniProt
  • Database links
  • Alternative names
    • CEPR antibody
    • Chemoattractant receptor-like 2 antibody
    • Chemokine receptor-like 2 antibody
    • CMKRL2 antibody
    • Constitutively expressed peptide like receptor antibody
    • DRY12 antibody
    • FEG 1 antibody
    • FEG-1 antibody
    • Flow-induced endothelial G protein-coupled receptor antibody
    • Flow-induced endothelial G-protein coupled receptor 1 antibody
    • G protein-coupled receptor 30 antibody
    • G-protein coupled estrogen receptor 1 antibody
    • G-protein coupled receptor 30 antibody
    • GPCR-BR antibody
    • Gper antibody
    • GPER_HUMAN antibody
    • GPER1 antibody
    • GPR30 antibody
    • IL8-related receptor DRY12 antibody
    • Lergu antibody
    • LERGU2 antibody
    • leucine rich protein in GPR30 3'UTR antibody
    • LYGPR antibody
    • Lymphocyte-derived G-protein coupled receptor antibody
    • Membrane estrogen receptor antibody
    • mER antibody
    • MGC99678 antibody
    see all

Images

  • Anti-G-protein coupled receptor 30 antibody (ab39742) at 1 µg/ml + Human brain normal tissue lysate - membrane extract (ab29456) at 10 µg

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

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 42 kDa
    Observed band size: 55 kDa
    why is the actual band size different from the predicted?



    GPR30 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.
  • ab39742 staining cultured rat primary astrocytes by ICC/IF. The cultured cells were fixed with 4% paraformaldehyde for 5 minutes and blocked with 10% donkey serum in 0.1% PBS-0.3% TritonX for 30 minutes at 24°C. The cultured cells were then stained with ab39742 at 1/500 in 0.3% TritonX with 0.1% PBS and 10% donkey serum for 4h at 24°C. An Alexa Fluro 488 donkey anti-rabbit polyclonal antibody at 1/1000 was used as the secondary antibody. Nuclei were stained with 1.43µM Hoechst and can be observed in blue. In red astrocytes can be observed (monoclonal anti GFAP).

    See Abreview

  • ICC/IF image of ab39742 stained U87-MG cells. The cells were 4% formaldehyde fixed (10 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 ab39742 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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.

  • Immunohistochemical analysis of Human ovarian endometriosis tissue, staining G-protein coupled receptor 30 with ab39742 at 1/50 dilution.

References

This product has been referenced in:
  • Gogola J  et al. Persistent endocrine-disrupting chemicals found in human follicular fluid stimulate the proliferation of granulosa tumor spheroids via GPR30 and IGF1R but not via the classic estrogen receptors. Chemosphere 217:100-110 (2019). Read more (PubMed: 30414542) »
  • Cortes E  et al. GPER is a mechanoregulator of pancreatic stellate cells and the tumor microenvironment. EMBO Rep 20:N/A (2019). Read more (PubMed: 30538117) »
See all 29 Publications for this product

Customer reviews and Q&As

1-9 of 9 Q&A

Answer

Thank you for contacting us.
I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.
In order to understand the problems encountered more fully and be able to offer any tips which may improve the results observed so far, would you mind answering the following questions:
1. Sample preparation: Lysis buffer: Protease inhibitors: Reducing agent: Heated prior to loading on gel? Temperature/time:
2. Was a loading control used?
3. Which secondary antibody is being used? Has this been used with any other primary antibody before? What have been the results of this?
4. Order number (or if you do not have access to this the approximate delivery date and the delivery address used).
This information will also allow us to investigate this case internally and initiate additional testing where necessary.
If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.
I look forward to receiving your reply.

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Answer

Thank you for contacting us. Can you tell me how your oocyte samples were fixed and embedded? Were you using fluorescent or colorimetric detection? Fluorescent signal can easily fade after storage for a few days, whereas colorimetric signal will be stable for months to years at room temperature, provided that the samples are properly fixed and embedded. How were you storing your samples? Were they mounted to a slide and coverslipped, or were they in solution? The signal should be more stable when the samples are mounted. I hope this helps, please let me know if you need any additional information or assistance.

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Answer

Thank you for contacting us.

My Spanish colleague is currently out of the office.

About the ER alpha R andER beta R antibodies:
The antibodies Ariana suggested to you are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

About the anti-Mas (https://www.abcam.com/ab123326 and https://www.abcam.com/ab66030) and the anti-GPER R (https://www.abcam.com/ab39742):
To our knowledge,these antibodieshave not been tested in IP.
Therefore, I can offer a discount off a future purchase if you buythese antibodiesnow, testthem inIP and submit feedback to us in the form of an Abreview.
It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback.
The discount would be to the value of 1 free primary antibody.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know which antibody you would like tobuy and test in IP. I will then send a discount code. This code must be issued before purchasingthe antibodies you want to test,so please wait for my reply before ordering.

2. Purchasethe antibody you want to testeither by phone, fax, or online (https://www.abcam.com).

3. Test it/them in IP.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for anyprimary antibodyordered and the discount code is valid for 4 months after issue.
Please remember that submission of the Abreview is sufficient for the discount code to become active.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even ifthe tested antibodyturns out to be unsuitable for IP, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: https://www.abcam.com/collaborationdiscount.

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Answer

Gracias por contactarnos.



He llevado a cabo una búsqueda en nuestro catálogo de anticuerpos contra los receptores indicados:



-Para MAS, tenemos dos opciones, pero lamentablemente ninguna de ellas se ha validado mediante IP, pero si en WB. Si queréis echar un vistazo a sus datasheets adjunto el enlace a ellas:



https://www.abcam.com/MAS1-antibody-ab123326.html



https://www.abcam.com/MAS1-antibody-ab66030.html



-Contra GPER, también hay dos anticuerpos disponibles, aunque me temo que tampoco se han usado en IP. Uno de ellos sin embargo se ha validado en WB:



https://www.abcam.com/G-protein-coupled-receptor-30-antibody-ab39742.html



-Contra ER alpha Receptor, hay varios anticuerpos validados mediante IP con células humanas, podéis echar un vistazo a las distintas alternativas en el siguiente enlace:



https://www.abcam.com/index.html?pageconfig=searchresults&search=ER%20alpha%20Receptor&pt=1&sk=app&sv=70&sn=IP&l=3&fViewMore=1



-Por último, para Estrogen Receptor beta, también hay dos alternativas validadas en IP con células humanas:



https://www.abcam.com/Estrogen-Receptor-beta-antibody-988-ab16813.html



https://www.abcam.com/Estrogen-Receptor-beta-antibody-EPR3777-ab92306.html






Siento que para los dos primeros receptores no haya una alternativa adecuada, sin embargo si queréis, podéis participar en nuestro sistema de evaluación de anticuerpos Abreview.

En Abcam ofrecemos un servicio de evaluación de anticuerpos, llamado Abreview, por el cual podemos ofrecerte un descuento (por valor de un anticuerpo primario / proteína) para una futura compra si adquieres el producto y lo testas en una aplicación o en una especie no testada hasta ahora (como IP), y publicas los resultados en nuestra website, en forma de Abreview, independientemente de que éstos sean positivos o negativos.

Si estás interesado en la oferta, házmelo saber antes de comprar los anticuerpos, para que te envie el código canjeable por un anticuerpo gratuito en la futura compra.

Para conocer con más detalle nuestro sistema de Abreview, visita la página: https://www.abcam.com/abreview.

No dudes en ponerte en contacto con nosotros si tienes cualquier otra consulta sobre esta oferta y estaré encantada de poder ayudarte. Si necesitas un presupuesto con alguno de los productos mencionados, también estaré encantada de enviártelo.

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Answer

Thank you for getting back to me regarding this case.



I have gone ahead and arranged for ab52650 Anti-PTP1B antibody [EP1837Y]to be shipped to you using the same purchasing route as before. We currently do not have local stock and our overseas stock will only reach you some time next week.Please also inform yourlocal sub-dealer (宮崎化学品株式会社) that you are expecting this free of charge product from us so that they too will be looking out for it. (Order # 1113473)



If there is anything else that I can help you with please do not hesitate to contact me.



Have a nice day!

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Answer

Thank you for your feedback.


I am sorry to hear that the different lot replacement vial also did not work as you hoped. Should you be interested in a different target primary antibody from our catalogue, I will be more than happy to offer you a free primary antibody of your choice. Simply inform me the catalogue number and I will arrange for the product to be shipped to you.


I hope this one incident will not hinderyou from considering us again as your source of antibodies and reagents in the future.


Thank you and good luck in your research.

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Question
Answer

Thank you for contacting us.  We currently have three antibodies that recognize GPCR GPR30: ab39742 - rabbit polyclonal; reacts with: human, mouse, and rat; tested for IHC-Fr and WB ab118512 - goat polyclonal; reacts with human and predicted to react with mouse and rat; tested for IHC-P ab98075 - rabbit polyclonal; reacts with human; tested for Flow Cyt, ICC/IF, and IHC-P. I hope this helps, please let me know if you need any additional information. 

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Answer

Thank you for your enquiry. I am sorry to hear that you are experiencing difficulties with this product ab39742 in western blot. Since you mentioned that you are getting expected results in immunohistochemistry but not in western blot (WB), the problem may be with the WB application. If you would instantly like a replacement with a different lot or product, I will be more than happy to arrange that for you. Please provide the name of your sub-dealer and sales person so that I can trace your original order number. If you would like some troubleshooting tips, here are some suggestions. 1) I can see from the image that the whole blot is rather dark and have streak of bands. Here at Abcam , we normally use 5% BSA to block membranes and do get cleaner images. If you have BSA in your lab, you may want to try it out. Also, performing a no-primary antibody experiment will be helpful in determining if your secondary antibody is causing the non-specific bands or not. 2) Although we normally recommend loading about 20-40ug of protein, this sometimes is more than enough and cases multiple bands and /or high background. If possible, could you try 10ug sample?  3) Lastly, boiling of sample is done at 98°C for 10 min to ensure that the protein is in the correct conformation to run at the correct molecular weight and be detected by the antibody. I hope the tips may be helpful. If not, then please do not hesitate to contact me. Also, please advise me on how you would like to proceed with your enquiry, so that I can immediately arrange for a replacement to you. I look forward to hearing from you again in order to resolve the matter.  

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Answer

Thank you for bringing this to our attention.

We have received several complaints over the past two years but without consistent observations, and none similar to yours. Our own data has the protein appearing at about 52 kDa. The assumption has been that glycosylations slow down migration in the gel. The predicted molecular weight of 42 kDa is based on the primary amino acid sequence, before modification post-translation.

So it is difficult to confidently say that the bands near 45 kDa in your blots are in fact GPR30 or are instead another protein that the antibody cross-reacts with. For the rat brain samples, a smear rather than two distinct bands is more expected, as a consequence of different amounts of glycosylation. We have not seen a band at 70 kDa before.

Can you please tell me how the samples were prepared before loading in to the gel? Also, how much protein was loaded per lane? Have you staining blots of these samples for any other proteins, and if so, were the results as expected?

Can you please send the order number or approximate date of the order? If I cannot make a suggestion, we have another lot available which may be worth a try.

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