Recombinant
RabMAb

Recombinant Anti-Muscarinic Acetylcholine Receptor 2/CM2 antibody [EPR4568] (ab109226)

Reviews (1)Q&A (8)References (7)

Overview

  • Product name

    Anti-Muscarinic Acetylcholine Receptor 2/CM2 antibody [EPR4568]
    See all Muscarinic Acetylcholine Receptor 2/CM2 primary antibodies

  • Description

    Rabbit monoclonal [EPR4568] to Muscarinic Acetylcholine Receptor 2/CM2

  • Host species

    Rabbit

  • Specificity

    Please note: blast analysis of the immunogen sequence for ab109226 shows 64% homology with the Muscarinic Acetylcholine Receptor 3, therefore some reactivity with this receptor subtype may be possible.

  • Tested applications

    Suitable for: WB, IP, IHC-Pmore details
    Unsuitable for: Flow Cyt

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human Muscarinic Acetylcholine Receptor 2/CM2 aa 150-250. The exact sequence is proprietary.
    Database link: P08172

  • Positive control

    • WB: SH-SY5Y, fetal brain, U87-MG, Neuro-2a, and Y79 lysates IHC-P: Human brain tissue IF: SH-SY5Y cells

  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab109226 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 1/1000 - 1/10000. Detects a band of approximately 52 kDa (predicted molecular weight: 52 kDa).
IP 1/10 - 1/100.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for Flow Cyt.

    • Target

    • Function

      The muscarinic acetylcholine receptor mediates various cellular responses, including inhibition of adenylate cyclase, breakdown of phosphoinositides and modulation of potassium channels through the action of G proteins. Primary transducing effect is adenylate cyclase inhibition.

    • Involvement in disease

      Genetic variations in CHRM2 can influence susceptibility to major depressive disorder (MDD) [MIM:608516]. MDD is one of the most common psychiatric disorders. MDD is a complex trait characterized by one or more major depressive episodes without a history of manic, mixed, or hypomanic episodes. A major depressive episode is characterized by at least 2 weeks during which there is a new onset or clear worsening of either depressed mood or loss of interest or pleasure in nearly all activities. Four additional symptoms must also be present including changes in appetite, weight, sleep, and psychomotor activity; decreased energy; feelings of worthlessness or guilt; difficulty thinking, concentrating, or making decisions; or recurrent thoughts of death or suicidal ideation, plans, or attempts. The episode must be accompanied by distress or impairment in social, occupational, or other important areas of functioning.

    • Sequence similarities

      Belongs to the G-protein coupled receptor 1 family. Muscarinic acetylcholine receptor subfamily. CHRM2 sub-subfamily.

    • Cellular localization

      Cell membrane. Cell junction > synapse > postsynaptic cell membrane.
    • Information by UniProt

    • Database links

      see all

    • Alternative names

      • 7TM receptor antibody
      • Acetylcholine receptor, muscarinic, 2 antibody
      • AChR antibody
      • Acm2 antibody
      • ACM2_HUMAN antibody
      • Cholinergic receptor muscarinic 2 antibody
      • Cholinergic receptor, muscarinic 2, cardiac antibody
      • Cholinergic receptor, muscarinic 2, isoform a antibody
      • Cholinergic receptor, muscarinic 2a antibody
      • CHRM 2 antibody
      • CHRM2 antibody
      • chrm2a antibody
      • CM2 antibody
      • FLJ43243 antibody
      • HM 2 antibody
      • HM2 antibody
      • M2 antibody
      • M2 muscarinic receptor antibody
      • M2-mAChR antibody
      • MGC120006 antibody
      • MGC120007 antibody
      • Muscarinic acetylcholine receptor M2 antibody
      • Muscarinic M2 receptor antibody
      see all

    Images

    • Western blot - Anti-Muscarinic Acetylcholine Receptor 2/CM2 antibody [EPR4568] (ab109226)
      Western blot - Anti-Muscarinic Acetylcholine Receptor 2/CM2 antibody [EPR4568] (ab109226)
      All lanes : Anti-Muscarinic Acetylcholine Receptor 2/CM2 antibody [EPR4568] (ab109226) at 1/1000 dilution

      Lane 1 : SH-SY5Y lysate
      Lane 2 : Fetal brain lysate
      Lane 3 : U87-MG lysate
      Lane 4 : Neuro-2a lysate
      Lane 5 : Y79 lysates

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Standard HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 52 kDa
      Observed band size: 52 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Muscarinic Acetylcholine Receptor 2/CM2 antibody [EPR4568] (ab109226)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Muscarinic Acetylcholine Receptor 2/CM2 antibody [EPR4568] (ab109226)

      Immunohistochemical analysis of paraffin-embedded Human brain tissue using ab109226 at 1/100.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    References

    This product has been referenced in:

    • Bachmann SB  et al. A Distinct Role of the Autonomic Nervous System in Modulating the Function of Lymphatic Vessels under Physiological and Tumor-Draining Conditions. Cell Rep 27:3305-3314.e13 (2019). Read more (PubMed: 31189113) »
    • Kashyap MP  et al. BDNF overexpression in the bladder induces neuronal changes to mediate bladder overactivity. Am J Physiol Renal Physiol 315:F45-F56 (2018). Read more (PubMed: 29092846) »
    See all 7 Publications for this product

    Publishing research using ab109226? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-9 of 9 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Muscarinic Acetylcholine Receptor 2/CM2 antibody [EPR4568]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Tendon)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Commercial High pH Buffer
    Permeabilization
    No
    Specification
    Tendon
    Blocking step
    Commercial Peroxidase blocking solution as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: RT°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 14 2014

    Question

    The customer would like refund.
    1005355
    PO#111221-1
    2013-1-14

    Read More
    Answer

    Thank you very much for your email.

    As we are still retesting this antibody so customer data would be very helpful. Could you please ask customer few questions;

    - cell line used
    - Problem; high background, no staining etc.
    - antibody dilution used
    - cell fixation procedure.


    Please send the details and we will process the refund.

    Many thanks for your cooperation in this case.

    Read More

    Question

    Thank you for your email. We have not finished investigating this matter. I will notify you as soon as I hear back from our QC team.

    I apologize for the delays and thank you for your understanding.

    Kind regards,

    Read More
    Answer

    Thanks for the update.

    I will look forward to hearing back soon.

    Read More

    Question

    I was wondering if you could please check form me what the delivery schedule for the replacement antibody is. We have not received it yet, and we would dearly need it for experiments!

    Thanks for your help!

    Read More
    Answer

    I apologize for the delay.

    The product will be delivered tomorrow morning. The order number is 123772 (ab32087).

    I hope this information will be helpful.

    Read More

    Question

    Thanks for your follow up. I'd prefer a refund for this product.
    Thanks,

    Read More
    Answer

    Thank you for contacting us.
    Your credit note ID is XXXXX.
    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.

    Read More

    Question

    thank you for the offer to replace the above mentioned antibody with another one. As I did not find any suitable M2R antibody in your catalog, I would like to exchange this one to a completely different one. Is this OK?

    What I would like to have is the antibody ab32087 (Anti MCL-1 antibody Y37)

    How should I proceed with this, do I need to order it or will you be taking care of the shipping just like this?

    Thanks a lot!

    Read More
    Answer

    I can send you the replacement, no problem!

    Could you confirm if you have received this antibody ab109226 as free of charge under the 100% Abreview testing promotion?

    Thanks!

    Read More

    Question

    1) Abcam product code ab 109226
    2) Abcam order reference number or product batch number GR50911
    3) Description of the problem Antibody: does not show expected membrane localization of muscarinic receptor, instead the staining resembles the pattern of microtubule and colocalizes with tubulin!
    4) Sample preparation:Species Homo SapiensType of sample: Cells in culture (HeLa cells)
    Sample preparation Cells were grown on coverslips with serum-containing mediumPositive control Negative control 5) Fixation step Yes If yes: Fixative agent and concentration Following conditions were tested for this antibody: Fixation in 100% methanol and in 4% paraformaldehyde.
    Fixation time 10 min Fixation temperature Fixation with methanol at -20°C; fixation with paraformaldehyde at room temperature 6) Antigen retrieval method 7) Permeabilization method:
    Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers? Permeabilizing agent and concentration: Cells fixed in 4% paraformaldehyde were permeabilized with 50 µg/ml digitonin. For MeOH, no permeabilization necessary.
    8) Blocking agent (eg BSA, serum…): Concentration 1% BSA in PBS Blocking time 10 min Blocking temperature room temperature 9) Endogenous peroxidases blocked? not relevant Endogenous biotins blocked? not relevant 10) Primary antibody (If more than one was used, describe in “additional notes”) : Concentration or dilution 1:100 Diluent buffer 1% BSA in PBS Incubation time 60 min
    11) Secondary antibody: Species: Donkey Reacts against: rabbit Concentration or dilution 1:300
    Diluent buffer 1% BSA in PBS Incubation time 60 min Fluorochrome Alexa-488, Cy3
    12) Washing after primary and secondary antibodies: Buffer PBS Number of washes 3 times in PBS and one quick wash in water 13) Detection method Immunofluorescence
    14) How many times have you run this staining? Twice Do you obtain the same results every time? Yes What steps have you altered to try and optimize the use of this antibody? Fixation also performed in paraformaldehyde. Document attachment: Attaching images of your IHC is strongly recommended and can greatly speed up our investigation of your problem.

    Read More
    Answer

    Thank you for your email and for providing the details.

    I have contacted our lab for the tubulin cross reactivity data. I will get back to you soon.

    Read More

    Question

    ich schreibe das auf Englisch, für den Fall, dass Sie es weiterschicken müssen

    We have recently ordered an antibody Ab109226 against muscarinic acetylcholine receptor 2 (Lot GR50911). This antibody has been declared to work in Western blot and immunocytochemistry. In WB, we got the expected band around 50 kDa plus some higher MW bands. However, we were not able to see any difference between nontransfected cells and those transfected with an M2R plasmid.

    We also tested this Ab in immunofluorescence, and the staining pattern does not correspond to the expected membrane protein pattern. Instead, we see what clearly looks like a tubulin cytoskeleton! Thus, we performed double staining with the M2R antibody (see enclosed picture M2R_HeLa_MeOH) and alpha-tubulin (Tubulin_HeLa_MeOH), and the stainings indeed overlap to a very large degree! In Addition, if you look at the picture M2R in mitotic cell, you will see that in a dividing cell, the staining is concentrated in the midbody region where you expect to see the tubulin cytoskeleton. This would not be the expected location of the M2R! Fixation with PFA produces very similar results, so this is not the cause…

    I was wondering if you have any idea what is wrong with this antibody? We purchased it since we were very happy to see that there is an antibody that works for several methods. Could it be that this specific lot contains a wrong antibody (labeled with wrong number?) instead of the M2R ab, since the staining that you have in your website indeed looks more like M2R. For WB, you cannot tell the difference since both proteins are close to 50 kDa.

    I would deeply appreciate if you could solve this mystery since we dearly need a functional antibody for this protein. If this turns out to be a lot-specific problem, we would be happy to receive a replacement of a batch that works! Otherwise, I would like to receive our money back since this Ab definitely does not recognize what it is supposed to.

    Thanks a lot!

    Read More
    Answer

    Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.

    I can confirm that, no other complaint was received for this antibody so the problem could be either with antibody or the protocol. We also do not mislabel products so the antibody you have received is indeed correct product.

    Multiple bands or high background problem is quite common with antibodies which can be overcome with troubleshooting so I am very much interested in checking the protocol you have been using. I have attached WB and IHC/ICC questionnaire for getting the information we need to make any decision regarding the quality of lot GR50911.

    Please fill these document and send these to me as per your earliest convenience.

    I will look forward to hearing from you soon.

    Read More

    Question

    Blocking peptide for ab2805

    Read More
    Answer

    Thank you for contacting us.

    Unfortunately we do not have blocking peptides for any of the antibody ab2805, ab90805 and ab109226. I apologize for the inconvenience this may have caused.

    I can however recommend trying the following protocol steps as alternative˜
    - use negative control cells or tissue that does not express this protein. The human protein Atlas database can help in finding this.
    - Use no primary control sections
    - Use isotype control antibody

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

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com