Overview

  • Product name
    Anti-Cannabinoid Receptor II antibody
    See all Cannabinoid Receptor II primary antibodies
  • Description
    Rabbit polyclonal to Cannabinoid Receptor II
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-Fr, WB, IHC-P, ICC, ICC/IF, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Chinese hamster
  • Immunogen

    Fusion protein corresponding to Rat Cannabinoid Receptor II aa 1-32 (N terminal).

  • Positive control
    • AtT20 cells.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.05% Sodium azide
    Constituents: 50% Glycerol, 0.1% BSA, 49% PBS
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Primary antibody notes
    Cannabinoids exert their well known physiological effects through two G protein coupled receptors, cannabinoid receptor 1 (CB1) and CB2. Both cannabinoid receptors have been shown to inhibit adenylyl cyclase as well as stimulate the mitogen-activated protein kinase, MAPK. CB1 receptors also modulate ion channels through direct G-protein interactions. Delta 9-tetrahydrocannibinol and related ligands likely exert their psychoactive effects by inhibiting presynaptic N- and P / Q type calcium channels. CB2 is thought to function primarily in the immune system although it has been suggested to be present in the central nervous system, including the retina.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab3561 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 1/500.
WB 1/50 - 1/500. Predicted molecular weight: 40 kDa.
IHC-P 1/10 - 1/100.
ICC 1/500.
ICC/IF 1/500 - 1/5000.
Flow Cyt Use at an assay dependent concentration. PubMed: 18178718

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

 

Target

  • Function
    Heterotrimeric G protein-coupled receptor for endocannabinoid 2-arachidonoylglycerol mediating inhibition of adenylate cyclase. May function in inflammatory response, nociceptive transmission and bone homeostasis.
  • Tissue specificity
    Preferentially expressed in cells of the immune system with higher expression in B cells and NK cells (at protein level). Expressed in skin in suprabasal layers and hair follicles (at protein level). Highly expressed in tonsil and to a lower extent in spleen, peripheral blood mononuclear cells, and thymus. PubMed:14657172 could not detect expression in normal brain. Expressed in brain by perivascular microglial cells and dorsal root glanglion sensory neurons (at protein level).
  • Sequence similarities
    Belongs to the G-protein coupled receptor 1 family.
  • Post-translational
    modifications
    Constitutively phosphorylated on Ser-352; phosphorylation increases cell internalization and desensitizes the receptor.
  • Cellular localization
    Cell membrane. Cell projection > dendrite. Perikaryon. Localizes to apical dendrite of pyramidal neurons.
  • Information by UniProt
  • Database links
  • Alternative names
    • Cannabinoid receptor 2 antibody
    • Cannabinoid receptor 2 macrophage antibody
    • CB 2 antibody
    • CB-2 antibody
    • CB2 antibody
    • CNR2 antibody
    • CNR2_HUMAN antibody
    • CNRII antibody
    • CX 5 antibody
    • CX5 antibody
    • hCB2 antibody
    • testis-dominant CNR2 isoform CB2 antibody
    see all

Images

  • ab3561 labelling Cannabinoid Receptor II in the cytoplasm of Human tonsil tissue (right) compared with a negative control (left) by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Tissues were blocked in 3% H2O2-methanol for 15 min at room temperature. Tssiue sections were incubated with the primary antibody (1:20 in 3% BSA-PBS) overnight at 4°C. A anti-rabbit HRP was used as the secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

  • ab3561 labelling Cannabinoid Receptor II in the cytoplasm of Human skin tissue (right) compared with a negative control (left) by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Tissues were blocked in 3% H2O2-methanol for 15 min at room temperature. Tssiue sections were incubated with the primary antibody (1:20 in 3% BSA-PBS) overnight at 4°C. A anti-rabbit HRP was used as the secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

  • All lanes : Anti-Cannabinoid Receptor II antibody (ab3561) at 1/200 dilution

    Lane 1 : HT29 cell lysate
    Lane 2 : C6 cell lysate
    Lane 3 : Rat colon cell lysate

    Lysates/proteins at 25 µg per lane.

    Predicted band size: 40 kDa
    Observed band size: 40 kDa

  • ab3561 staining Cannabinoid Receptor II in murine BV2 cells by Immunocytochemistry/ Immunofluorescence. Cells were stimulated with 10 uM ADP for 15 minutes at 37°C, 5%CO2 then washed 3X with PBS and fixed with 2% formaldehyde for 15 minutes at room temperature. A blocking step was performed for 1 hour at room temperature with 5% FBS/PBS/0.01% Triton-X-100. ab3561 used at a 1/500 dilution for 1 hour at room temperature then washed 3X PBS. The secondary was an Alexa Fluor 488 conjugated antibody, used at a 1/250 for 1 hour at room temperature, washed 3X PBS then mounted in Vectashield containing DAPI.
  • Quantification of CB1 and CB2 receptor surface expression in primary Human and HaCaT keratinocytes. (A) FL-1 shows autofluorescence (black population), non-specific binding of secondary antibody (dark grey population) and specific immunofluorescence obtained with CB1 (ab3558) and CB2 (ab3561) receptor antibodies, respectively (light grey population). Histogram shows representative measurement (5000 cells counted). (B) Differences in CB receptor expression between primary and HaCaT keratinocytes, showing marked increased expression of CB2 in primary keratinocytes (geo mean corrected for non-specific binding of secondary Ab). Data show mean values ±SEM obtained with two human primary keratinocyte samples and HaCaT cells, each measured three times.

  • ab3561 at 1/5000 staining Rat spinal cord tissue sections by IHC-P. Following nerve trans-section, the animal was transcardially perfused with PBS followed by 4% formaldehyde (no post fixation) then frozen in OCT and stored at -80°C until required. 20µm sections were cut and stained with the antibody for 18 hours. A biotinylated goat anti-rabbit antibody was used as the secondary.

    See Abreview

  • Immunocytochemistry/immunofluorescence analysis of AtT20 cells transfect with the rat CB2 gene labeling Cannabinoid Receptor II with ab3561.

References

This product has been referenced in:
  • Wang B  et al. Restoration of osteogenic differentiation by overexpression of cannabinoid receptor 2 in bone marrow mesenchymal stem cells isolated from osteoporotic patients. Exp Ther Med 15:357-364 (2018). Read more (PubMed: 29250156) »
  • 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 26 Publications for this product

Customer reviews and Q&As

1-10 of 31 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HT29 cell line (colon cancer epithelial cell line))
Permeabilization
Yes - Tween 0,25%
Specification
HT29 cell line (colon cancer epithelial cell line)
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 23°C
Fixative
Methanol

Abcam user community

Verified customer

Submitted Oct 30 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (bladder)
Loading amount
100 µg
Specification
bladder
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (10% gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C

Dr. Eun-Young Park

Verified customer

Submitted Apr 09 2013

Answer

Thank you for that clarification.

I have sent you a FedEx link that you can complete to be able to return the vial of ab3561 from order number *. Once that order is returned to us, we will credit the cost of that product back to you. The credit note for this is at the bottom of the email.

As for the vial that you recieved on order number *, as we discussed I am sending you a vial of ab3560. The order number for this is 1* and you should have it on Tueday morning.

If there is anything else I can help you with, please let me know.

Credit Note for order #*

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. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

The credit note ID is for your reference only and does not automatically guarantee the credit.

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 expert advice.

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Question
Answer

Thank you for that information.

That order number (*), is for the vial of ab3561 that you just received today, correct? If so, I will go ahead and set up the return and credit for this order.

Do you have the order number for the first vial of ab3561 that you received, as I will need that number to send the free of charge replacement.

Sorry, I do not mean to make this complicated; I just want to make sure I replace/refund the correct orders.

Thank you for your patience and cooperation and I hope to have this resolved quickly.

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Answer

Thank you for your reply.

I am sorry about the frustration you are having with ab3561 in flow cytometry and I would be happy to replace the first vial that you received. Of the two possible replacements you mentioned in you previous email, we only guarantee ab3560 to work in flow cytometry, although we have only tested it in rat and human samples. I know you were transfecting CHO cells, but is the construct for rat or human?

For the vial of ab3561 that you just received, if you could please provide the order number for this product, I will make arrangements for the cost to be credited back to your account and also for you to return that vial to Abcam.

I look forward to your reply and helping to resolve this issue.

Read More

Answer

Thank you for contacting Abcam.

The antibody is covered under our Abpromise for six months and is guaranteed to work in flow cytometry on a number of samples. If we cannot resolve the issue you are having with the antibody then I would be happy to either send a replacement antibody or to process a refund.

The information we have regarding concentrations for the antibody working in flow cytometry, come from some references, which are available on the website. That information is also below:

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=20206138&dopt=Abstract - They used 1/100 dilution.
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=18239598&dopt=Abstract - They do not specify the concentration they used.



If you cannot find any protocol information in these references that prove to be useful, then please feel free to send me a copy of the protocol and results that you are getting and I would be happy to look them over and see if there is anything that I can suggest.

Please let me know if there is anything else I can help you with.

Read More

Answer

Thank you for contacting us.

The number of slides that could be used with this kit depends on the tissue sections, and the volume of reagents per slide. The volume used should be enough to cover them. We normally use 200ul per slide in house, but 100ul should be enough. If 100ul are used then you could run 150 slides with the reagents provided.

This kit is compatible with any primary antibody suitable for IHC in paraffin sections, as long as the recommendations and protocols are followed.

Regarding the tissue slides to use as controls, I checked the http://www.proteinatlas.org/ENSG00000188822, which suggests digestive tract, female tissues or kidney as positive controls. Other tissues could be Human Spinal Cord, Rat Spinal Cord, and Mouse Thymus. I can check if you are interested what would be a good negative control with my lab colleagues.

If you are using the EXPOSE kit there is no need to use an enhancer for detection.

Except for the products ab94726 and ab64026, the rest of them are currently in stock, and you will receive them within two working days.

Please note all of our products are covered by our Abpromise® guarantee, which ensures that you can trust our products, and they should work in the tested species and applications stated on the datasheet, or we will offer a replacement, credit, or refund, if reported within 6 months of purchase.

To read about our Abpromise policy: https://www.abcam.com/index.html?pageconfig=abpromise

You will receive the proforma invoice with the requested products shortly.

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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Answer

Thank you very much for your reply and for sending the additional information.

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 at mailto:us.credits@abcam.com so that we may gather the correct information needed for the refund. 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.

Read More

Question
Answer

Thank you very much for your call today and for letting us know about the trouble with this antibody.

If you could please provide the following information I will arrange the refund promptly:

1) What kind of mouse samples were used (tissue or cell type)?
2) What application was the antibody used in?
3) Could you describe the results that were seen?
4) How were the samples prepared or fixed?
5) What kind of blocking solution was used?
6) At what dilution was the antibody used?


I look forward to hearing from you. Please let me know if you have any further questions or if there is anything else that we can do for you.

Read More

Answer

Thank you for contacting us.

We have following CB1 and CB2 available. These are testes in flow cytometry and are fully guaranteed.

ab3558; https://www.abcam.com/Cannabinoid-Receptor-I-antibody-ab3558.html

ab3561; https://www.abcam.com/Cannabinoid-Receptor-II-antibody-ab3561.html

ab3560; https://www.abcam.com/Cannabinoid-Receptor-II-antibody-ab3560.html

Could you let me know which conjugated you would like for compatible secondary antibody?

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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1-10 of 31 Abreviews or Q&A

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