Overview

  • Product name
    Anti-Mu Opioid Receptor antibody
    See all Mu Opioid Receptor primary antibodies
  • Description
    Rabbit polyclonal to Mu Opioid Receptor
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-Fr, ICC, IHC-FoFrmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Guinea pig, Human
    Predicted to work with: Cow, Pig, Non human primates, Macaque monkey
  • Immunogen

    Synthetic peptide:

    NHQLENLEAETAPLP

    , corresponding to C terminal amino acids 384-398 of Rat Mu Opioid Receptor 1.

Properties

Applications

Our Abpromise guarantee covers the use of ab10275 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 1/100 - 1/2000.
WB 1/500 - 1/1000. Detects a band of approximately 49-65 kDa (predicted molecular weight: 45 kDa).Can be blocked with Rat Mu Opioid Receptor peptide (ab46988).

Approximately 49 -65 kDa depending on the tissue lysate. Can be blocked with ab46988.

IHC-Fr 1/100 - 1/2000. We recommend incubating for 18-24 hours at 4C.
ICC 1/100 - 1/2000.
IHC-FoFr 1/100 - 1/2000. We recommend incubating for 18-24 hours at 4C.

Target

Images

  • Ab10275 (1:2500) staining of Mu opioid receptor by IHC.
  • ICC/IF image of ab10275 stained SHSY5Y 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 (ab10275, 1 in 200 dilution) 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.

  • Immunohistochemical analysis of rat brain tissue, staining Mu Opioid Receptor with ab10275.

    Tissue was blocked with 5% normal donkey serum and 0.4% Triton X-100 for 1 hour at room temperature. Sections were incubated with primary antibody (1/2000) for 24 hours at 4°C. An AlexaFluor®594-conjugated donkey anti-rabbit IgG (1/250) was used as the secondary antibody.
  • Mu Opioid Receptor (ab10275) staining of Rat DRG (dilution: 1:100) incubation at 4 °C overnight. Secondary antibody is anti-Rabbit Rhodamine Red (dilution:1:200) incubation at room temperature for 1 hour.

References

This product has been referenced in:
  • Hou X  et al. Suppression of HDAC2 in Spinal Cord Alleviates Mechanical Hyperalgesia and Restores KCC2 Expression in a Rat Model of Bone Cancer Pain. Neuroscience 377:138-149 (2018). WB ; Rat . Read more (PubMed: 29482000) »
  • Wu ZY  et al. Endomorphin-2 Decreases Excitatory Synaptic Transmission in the Spinal Ventral Horn of the Rat. Front Neural Circuits 11:55 (2017). Read more (PubMed: 28848403) »
See all 25 Publications for this product

Customer reviews and Q&As

1-10 of 27 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Mouse, whole brain sections)
Antigen retrieval step
None
Permeabilization
Yes - 10% Tween-20
Specification
Mouse, whole brain sections
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 14 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - other (Gross brain dissected tissue)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
Gross brain dissected tissue
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Jul 26 2016

Answer

Gracias por contactarnos.
Después de consultar este caso con mis compañeros del laboratorio, acerca de los siguientes anticuerpos contra Mu Opioid Receptor validados en tejidos de rata: ab10275, ab17934, ab64746, ab128013 y ab134054, me temo que ninguno de ellos ha demostrado reconocer el epítopo extracelular de la proteína.
Todos ellos se han generado a partir del extremo C terminal de la proteína, por lo que, a pesar de no haberse mapeado el epítopo que reconocen, probablemente detecten el dominio citoplásmico de la misma.
La solución que te propongo es testar uno de los anticuerpos que hay en el catálogo hecho a partir del extremo N terminal de la proteína, y testarlo en rata. Tenemos en abcam un sistema de evaluación de anticuerpos por el cual al testar uno de ellos en una aplicación o especie que no se ha testado, puedes beneficiarte de un descuento en tu próxima compra por valor del anticuerpo, si lo usas en esa especie o aplicación nueva y nos mandas los resultados a la web en forma de Abreview.
Para saber más sobre este sistema: www.abcam.com/abtrial
De las posibles alternativas, ab140911, ab137460 o ab137277 indícame si quieres cuál os interesaría más, para averiguar su homología con la proteína en rata y determinar la posibilidad de reaccionar en esta especie, en caso de que queráis participar en el sistema Abtrial.
Espero haberte ayudado. No dudes en contactarnos de nuevo para cualquier otra consulta.

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Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxxxx.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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Question

Dear abcam,

1- Yes, it was an immunohistochemistry on perfusion-fixed samples. The animals were perfused in 4%PFA, and the brains were cut with a cryostat at 30 µm thickness.

2- I send you the images in a pdf file

3- I applied also a less strong antigen retrieval, 20 min 80°C 50mM NaCitrate pH 8.5. The slices treated this way didn’t show a strong staining where myelin is (as did the stronger antigen retrieval method), but still the staining was aspecific, and the slice, processed with DAB, became dark in 10-15 seconds, really faster than what it does usually. I think it might be due to the primary antibody binding aspecifically. I tried this antigen retrieval procedure because I contacted the authors of 2 of the papers you cited in the antibody page, and they sent me that protocol (even if they didn’t write it in the paper). I contacted them just because I tried without antigen retrieval steps, as I already mentioned in my past emails, but without success.

4- I used TritonX because the articles your page cites as reference for the antibody use TritonX, and at concentrations similar or higher than the ones I used.

5- All washing steps have been done in PBS pH 7.4, 3 washes of 15 minutes before and after each step (antigen retrieval, primary and secondary antibodies).

6- I didn’t perform a no primary control because I processed other slices with another antibody I know, and which works, and the same secondary antibody worked perfectly.

Since I am really in a hurry, and I need to perform these experiments in a short time, would you please send me another vial of the antibody from another lot?

Thank you very much,

Sincerely,

Read More
Answer

Thank you for your answer.

I am sorry to confirm that we don't have any different lots in stock at the moment. I would like to reassure you that we monitor feedback closely on a weekly basis and we are not currently concerned about the general quality of this antibody or this batch. Regrettably, I can suggest you have received a bad vial on this occasion.

Therefore, we would be pleased to provide a replacement from the same lot, which will still be covered by our guarantee or even a different antibody. Alternatively, I fully understand your concerns and if you prefer a credit note in this case, I will be pleased to arrange this for you.

I understand that your time is restricted and I appreciate the time you already spent in the laboratory. It would be beneficial, if it is possible for you to try the suggestions we made. We experienced that individual experiments might need different optimisation.

I hope this will be helpful. I look forward to hearing from you with details of how you would like to proceed.

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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 antibody.

The details you have kindly provided will enable us to investigate this case for you and also gives us 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. I would like to reassure you that this antibody is tested and covered by our guarantee for mice and ICC/IF, WB, IHC-Fr, ICC as well as IHC-FoFr applications.

Before deciding how to proceed with this particular case, I would like to offer some suggestions to help optimise the results. I would also appreciate if you can confirm some details of the procedure.

1. Could you please confirm what applications is performed? Was it an IHC-FoFr experiment - perfusion fixed samples?

2. Regrettably we cannot open the send images. Could you kindly provide images in a PDF format please?

3. The described antigen retrieval seems to be long. Because the Mu Opioid Receptor is a membrane protein we would recommend to optimise the antigen retrieval. We would suggest trying no antigen retrieval as well as 2, 5, 10 and 20 minutes.
I can confirm that reviewing the literature in comparable experiments no antigen retrieval was performed (Chuhma N et al. Functional connectome of the striatal medium spiny neuron. J Neurosci 31:1183-92 (2011). IHC-FoFr; Mouse.http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=21273403&dopt=Abstract)

4. Furthermore it might be beneficial to use a less strong detergent then TritonX in this case. TritonX might disrupt the membrane and by doing so prevent the detection of the Mu Opioid Receptor in the membrane. A more gentle detergent is for example Tween. Generally permeabilisation is not always required for mambrane proteins.

5. Could you confirm if and what kind of washing steps were performed in this experiment. Unspecific staining might be due to not suitable washing buffer. Washing steps need to be optimised for every experiment to achieve clear results.

6. We suggest that a no primary control might be beneficial to confirm if the secondary antibody binds unspecific and gives the unspecific staining in the samples.

We are happy to offer this technical support. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

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Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from our Anti-Mu Opioid Receptor antibody (ab10275).

I would like to reassure you that this antibody is tested and covered by our 6 month guarantee for applications and species cited on the data sheet. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily. I send the questionnaire for IHC. If you were performing WB please let me know so that I can send you the right questionnaire for the application.

It would be very useful for our quality control if you could provide detailed information about the optimisation steps that were performed.

I would appreciate if you could also provide an image which would help us to assess the results.

Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.

Order Details
Antibody code:

Lot number:

Purchase order number
or preferably Abcam order number:


General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, low signal, non-specific satining etc.)

Sample (Species/Tissue/Cell Type/Cell Line etc.)


Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.)


Antigen retrieval (Enzymatic method, Heat mediated technique etc.)


Permeabilization step


Blocking conditions (Buffer/time period, Blocking agent etc.)


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Detection method


Positive and negative controls used (please specify)


Optimization attempts (problem solving)
How many times have you tried the IHC?



Have you run a "No Primary" control?
Yes No

Do you obtain the same results every time?
Yes No


What steps have you altered?


Additional Notes


We would appreciate if you are also able to provide and image which woudl help us to assess the results

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Question
Answer

Thank you for contacting us and letting us know about the trouble with this antibody.

Your credit note ID is ***

I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. Our accounting department can be contacted by email at us.credits@abcam.com or by telephone at 888-772-2226. Please refer to the credit note ID in any correspondence with our accounting department.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further assistance or advice.

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Answer

Thanks for your enquiry.

ab10275 is a whole antiserum product and the exact concentration of the anti-Mu opioid receptor in the serum has not been determined. In general, however, specific antibody concentration inproducts of this type is typically 0.5mg/ml or less.

I hope this is helpful. Please contact us again if you have any further questions.

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1-10 of 27 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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