Overview

  • Product name
    Anti-Muscarinic Acetylcholine Receptor 2 antibody [31-1D1]
    See all Muscarinic Acetylcholine Receptor 2 primary antibodies
  • Description
    Mouse monoclonal [31-1D1] to Muscarinic Acetylcholine Receptor 2
  • Host species
    Mouse
  • Specificity
    This antibody is specific for the m2 mAChR subtype.
  • Tested applications
    Suitable for: WB, Flow Cyt, ICC/IF, ELISA, IHC-P, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig
    Does not react with: Chicken
  • Immunogen

    Full length native protein (purified) corresponding to Pig Muscarinic Acetylcholine Receptor 2. Purified from Pig Heart.

Properties

Applications

Our Abpromise guarantee covers the use of ab2805 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. Detects a band of approximately 64 kDa (predicted molecular weight: 52 kDa).
Flow Cyt Use at an assay dependent concentration.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

ICC/IF Use a concentration of 5 µg/ml. PubMed: 19889856
ELISA Use at an assay dependent concentration.
IHC-P 1/20.
IP Use at an assay dependent concentration.

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
  • 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

  • Immunohistochemistry was performed on normal biopsies of deparaffinized human kidney tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer and microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a Mouse monoclonal antibody recognizing Muscarinic Acetylcholine Receptor 2 (ab2805) or without primary antibody (negative control) overnight at 4�C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
  • ICC/IF image of ab2805 stained PC12 cells. The cells were 4% PFA 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 (ab2805, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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.

  • All lanes : Anti-Muscarinic Acetylcholine Receptor 2 antibody [31-1D1] (ab2805) at 1 µg/ml

    Lane 1 : Human brain tissue lysate - total protein (ab29466)
    Lane 2 : Human spinal cord tissue lysate - total protein (ab29188)
    Lane 3 : Brain (Mouse) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 52 kDa
    Observed band size: 64 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 22 kDa, 40 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 30 seconds


    Muscarinic Acetylcholine Receptor 2 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

References

This product has been referenced in:
  • Benoit A  et al. Flow cytometry for receptor analysis from ex-vivo brain tissue in adult rat. J Neurosci Methods 304:11-23 (2018). Flow Cyt . Read more (PubMed: 29660368) »
  • Jovanovic P  et al. Oxytocin modulates the expression of norepinephrine transporter, ß3-adrenoceptors and muscarinic M2 receptors in the hearts of socially isolated rats. Peptides N/A:N/A (2018). Read more (PubMed: 29969648) »
See all 8 Publications for this product

Customer reviews and Q&As

1-10 of 12 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (40 ´m skin sections)
Permeabilization
Yes - Triton X
Specification
40 ´m skin sections
Blocking step
BSA as blocking agent for 30 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Paraformaldehyde

Mr. Christoph Erbacher

Verified customer

Submitted Aug 30 2017

Application
Western blot
Sample
Rat Tissue lysate - whole (Intestinal tissue)
Gel Running Conditions
Reduced Denaturing (4-12% BIS TRIS)
Loading amount
15 µg
Specification
Intestinal tissue
Blocking step
Milk as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Jul 13 2016

Application
Western blot
Sample
Mouse Tissue lysate - whole (Intestinal tissue)
Gel Running Conditions
Reduced Denaturing
Loading amount
14 µg
Specification
Intestinal tissue
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Mr. Julien Chevalier

Verified customer

Submitted Jul 08 2016

Application
Western blot
Sample
Human Tissue lysate - whole (Intestinal tissue)
Gel Running Conditions
Reduced Denaturing
Loading amount
14 µg
Specification
Intestinal tissue
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 08 2016

Application
Western blot
Sample
Pig Tissue lysate - whole (Intestinal tissue)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
Intestinal tissue
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 08 2016

Question
Answer

Thank you for contacting us. I hope that the suggestions which were made help you to resolve the issues which you have been experiencing. As requested, I have arranged for you to receive positive control lysates ab71719 and ab29330 free of charge. The confirmation number for these products is xxxxx. I have also created enrolled you into our 100% Abreview program for testing ab60981 in western blot. After submission of an Abreview the code xxxx will activate, entitling you to one free primary antibody of your choice. Further information about this program may be found at the following link:https://www.abcam.com/index.html?pageconfig=resource&rid=11998 I have also created for you a 25% off one item discount code which may be redeemed anytime before 17 January 2013. This 25% off code is xxxxx 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

Read More

Question
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

Question
Answer

Thank you for your enquiry. Unfortunately, we do not have the exact information you require but I was able to locate the following information in the system. 1) No cross-reaction with thodopsin which is highly homologous to mAChR by Western Blot. 2) Does not cross-react with M1 the neurological form of the receptor in IP (M1 and M2 are ~ 38% homologous). However, in checking the literature, the following article includes data that demonstrates the specificity of this antibody for M2 when tested against M1 - M4. P. Wang, G.R. Luthin and M.R. Ruggieri (1995). Muscarinic acetylcholine receptor subtypes mediating urinary bladder contractility and coupling to GTP binding proteins. J. Phamacol. and Exp. Therapeutics. 273 (2), 959 -273. I hope the above information will still be helpful.

Read More

Answer

The monoclonal antibody to the Muscarinic 2 Acetylcholine Receptor has not been mapped, therefore the epitope could be contained anywhere on the protein. The immunogen is purified muscarinic receptor from pig heart. This product is specific for M2 Receptor. I hope this information helps. Please do not hesitate to contact us if you need anything further.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell
Permeabilization
No

Abcam user community

Verified customer

Submitted Nov 02 2005

1-10 of 12 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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