Anti-Cannabinoid Receptor I antibody (ab23703)

Rabbit polyclonal Cannabinoid Receptor I antibody. Validated in WB, IHC, ICC/IF and tested in Mouse, Rat, Dog, Human. Cited in 36 publication(s). Independently reviewed in 17 review(s).

Overview

  • Product name
    Anti-Cannabinoid Receptor I antibody
    See all Cannabinoid Receptor I primary antibodies
  • Description
    Rabbit polyclonal to Cannabinoid Receptor I
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, IHC-Fr, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Dog, Human
    Predicted to work with: Cow, Chimpanzee, Macaque monkey
  • Immunogen

    Synthetic peptide:

    MSVSTDTSAE AL

    , corresponding to C terminal amino acids 461-472 of Human Cannabinoid Receptor I(Peptide available as ab50542.)

  • Positive control
    • Human cerebellum or murine brain sections.

Properties

Applications

Our Abpromise guarantee covers the use of ab23703 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
WB 1/200. Detects a band of approximately 60 kDa (predicted molecular weight: 53 kDa).Can be blocked with Cannabinoid Receptor I peptide (ab50542).
IHC-P Use at an assay dependent concentration.
IHC-Fr 1/300. Detection with an ABC system is recommended.
ICC/IF 1/50.

Target

Images

  • Anti-Cannabinoid Receptor I antibody (ab23703) + Human cerebellum at 30 µg

    Predicted band size: 53 kDa
    Observed band size: 60 kDa
    why is the actual band size different from the predicted?



    Secondary antibody - anti-rabbit HRP (ab6721)

  • Immunoperoxidase staining of CA region of rat hippocampus with the Cannabinoid Receptor I antibody (ab23703, 4ug/ml). the majority of the protein is localized in the pre-synaptic axons, shown as brown particulaes. A few neurons also show cytoplasm staining.
  • ab23703 staining Cannabinoid Receptor I in rat brain tissue by Immunohistochemistry (Frozen sections). After fixation in 4% PFA in PBS (pH 7.4), brains were cryoprotected in 30% sucrose in PBS at +4°C, frozen, and thereafter kept at -80°C until used. For the immunostaining, brains were cryosectioned in 30 µm slices, collected in PBS (pH 7.4), and processed in a free-floating system. Slices were first incubated in the blocking solution (BS) containing 2% bovine serum albumin, 2% goat serum, and 0.1% Triton X-100 in PBS (pH 7.4) for 1 hour at room temperature, and thereafter with the primary antibody for 24–48 hours at +4°C in BS at a 1/1000 dilution. After washing in PBS, slices were incubated with the biotin-conjugated secondary antibody (1/4000) in BS, rinsed with PBS, and incubated with the avidin-peroxidase conjugate in BS for 1 hour at room temperature. The staining was detected using 3,3-diaminobenzidinetetrahydrochloride as a chromogen.
  • Immunofluorescence analysis of mouse cultured hippocampal neurons, staining Cannabinoid Receptor I with ab23703.

    Cells were fixed with paraformaldehyde, permeabilized with 0.05% Saponin and blocked with 10% fetal calf serum for 2 hous at 4°C. Samples were incubated with primary antibody (1/200 in 0.05% Saponin + fetal calf serum) for 12 hours at 23°C. An AlexaFluor®488-conjugated goat anti-rabbit polyclonal IgG (1/400) was used as the secondary antibody.

    See Abreview

  • ab23703 staining Cannabinoid Receptor I in human pancreas tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at room temperature; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/50 in PBS) for 8 hours at 4°C. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Miragliotta V  et al. Cannabinoid receptor types 1 and 2 and peroxisome proliferator-activated receptor-a: distribution in the skin of clinically healthy cats and cats with hypersensitivity dermatitis. Vet Dermatol N/A:N/A (2018). Read more (PubMed: 29920828) »
  • Corrado A  et al. Endocannabinoid receptor CB2R is significantly expressed in aspirin-exacerbated respiratory disease: a pilot study. Int Forum Allergy Rhinol N/A:N/A (2018). Read more (PubMed: 29897665) »
See all 36 Publications for this product

Customer reviews and Q&As

11-20 of 32 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (mouse brain sections)
Specification
mouse brain sections
Fixative
Paraformaldehyde
Antigen retrieval step
None
Permeabilization
Yes - Triton during block and staining
Blocking step
PBS, 10%Horse serum, 0,2%BSA, 0,3%Triton as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Jan 07 2013

Answer

Thank you for contacting Abcam.

Earlier we discussed your use of ab23703 for IHC-P, specifically the lack of a positive signal with a 72oC Citrate buffer antigen retrieval. We discussed some troubleshooting tips, specifically attempting a 30 minute Citrate buffer antigen retrieval at 95oC water bath in a coplin jar.

Alternatively, enzymatic antigen retrieval may be preferential for the Cannabinoid Receptor 1. We discussed proteinase K and pepsin as potential enzymatic antigen retrieval buffers. Below is a link to a good resource for antigen retrieval buffers, including how to create the pepsin and proteinase K buffers:
http://www.ihcworld.com/epitope_retrieval.htm

Increasing your primary antibody concentration is another tip that may help. Below are further troubleshooting tips:
https://www.abcam.com/ps/pdf/protocols/ihc_p.pdf

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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Answer

Thank you for contacting us.

I am sorry to hear you have been experiencing problems with this antibody. The quality of our products is important to us and I would like to reassure you that we investigate all customer complaints.

I would like to investigate this particular case further for you, and also obtain more information for our quality records.

As far as I can understand, the problem you are experiencing with this antibody is high background? If not, please send us further information about what it is exactly.

Could you please provide the old lot number which gave successful results in the past? I would like to monitor the quality of this product and check whether there has been any issue with the current lot. For that, I would also need the order number for the last unit you received.

Have you used the same samples with this new antibody? Have there been any changes such as lysis buffer, enzymatic inhibitors, reducing agents, blocking conditions, etc…?

I look forward to receiving your reply to be able to help you solve this issue.

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Answer

Thank you for your reply and for providing the details of your protocol. There can be batch variability with polyclonal antibodies such as ab23703, and some batches may be stronger than others. All of our stock has passed our QC standards for specificity, and should produce good staining with a dilution of around 1:50 and an overnight incubation with the primary antibody. All of our antibodies will be guaranteed for 6 months from the date of purchase but should be stable for at least a year when stored according to the datasheet instructions. If the signal is weak, you could also try lengthening your antigen retrieval (we recommend Citrate buffer at pH 6.0).

I hope this helps to improve your results, if not, please let me know your original order or PO number and I will be happy to help you further.

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Answer

Thank you for contacting us.

The predicted molecular weight of ab23703 is 53KDa so it is normal for the customer to see a band running at 53KDa. The 60KDa band could be due to the post translationally modified form of the target protein. The targe protein got three glycosylation sites which could increase the molecular weight of the observed band.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Answer

DISCOUNT CODE: 42128 for ab3561 for WB image on Rat samples,

Expiration date: 27/10/2012



DISCOUNT CODE: 41516 for ab23703 for WB image on Rat samples,


Expiration date:27/10/2012


I am very pleased to hear you would like to use abxxxx in [SPECIES/APP] and send us an image of your results.


The code will be active once the Abreview with the image has been submitted and can be redeemed in one of the following ways:
1) Call to place your order and mention the code to our customer service department;
2) Include the code in your fax order;
3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated. Remember if the product does not work you will be covered by our Abpromise guarantee and eligible for a full refund or replacement.

If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Answer

Thank you for your response.

Thank you for your enquiry. In future, it would be much appreciated if discount code is requested before purchase. Please explain your customers that discount code can be requested for testing our products 1) in non-tested species or application or 2) in tested application when image is not available on the datasheet.

Could you please pass the customer's contact details for me so that I can issue the discount codes for ab3561 and ab23703 for the image.

I look forward to hearing from you soon.

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Answer

Thank you very much for your interest in ab3561 and ab23703.

I can confirm that both antibodies have been tested in Western blot applications and detect rat proteins; however we do not currently have an image of available for these two antibodies (WB in rat). Nevertheless, our Abpromise applies for these two antibodies.

Therefore, I can offer a discount off a future purchase if you buy either ab3561or ab23703 now, use it WB in rat and submit an image of your results to us in the form of an Abreview. The discount would be to the value of 1 free PRIMARY ANTIBODY.

It doesn’t matter whether the result is positive or negative, we would just really like to receive your feedback. Of course, if the results are negative you will be covered by our Abpromise guarantee and eligible for a refund or replacement.

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

1. Reply to this e-mail to let me know that you would like to proceed and send an image ofthe selected antibodyin WB in rat. I will then send a discount code. This code must be issued before purchasing the antibody so please wait for my reply before ordering.

2. Purchase the selected antibody either by phone, fax, or online (www.abcam.com).

3. Use it in WB in rat.

4. Send us an image of 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 any PRIMARY ANTIBODY ordered 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!

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: www.abcam.com/collaborationdiscount.

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Answer

Thank you for these additional details.
I would like to suggest the following steps:
1) Use another blocking solution to determine whether milk is affecting the antibody. We recommend using 3% BSA.
2) Try using higher dilutions of ab23703, 1:500 or even 1:100.
3) Try heating your samples for a longer period. If you are reluctant at 95C try 60C for 15 minutes as you have suggested.
Please let me know if these suggestions help solve this issue. This product is covered by our Abpromise guarantee. If we cannot remedy this issue and this is a product that you have purchased within the last six months, we will replace or refund it under our Abpromise guarantee, as you are using it according to specifications listed on our datasheet.
I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.
Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Answer

Thank you for contacting Abcam and for sending this image and details to me.
I am sorry to hear of the difficulties that you have been experiencing using these products. While this data has been enlightening, would you mind sending me some additional details of your protocol? Details such as blocking procedures, antibody incubation steps and concentrations which you have tried, what wash buffer have you used and any steps that you may have changed while troubleshooting would be helpful as well. These details will not only help me to better understand your difficulties and be able to offer more suggestions but will also be used for internal testing of the products if deemed necessary. If we are unable to solve the problems you are experiencing we will replace or refund the antibody according to our Abpromise guarantee.

Please let me know if you have any questions.

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11-20 of 32 Abreviews or Q&A

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