Immunofluorescence analysis of Muscarinic Acetylcholine Receptor M2 was done on 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled Muscarinic Acetylcholine Receptor M2 (31-1D1) Mouse Monoclonal Antibody (MA3044) at 2ug/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Pig, Rat|
|Published species reactivity||Rat, Pig, Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Purified porcine cardiac m2 muscarinic acetylcholine receptor.|
|Storage buffer||1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||Assay Dependent|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1:1,000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 3 publications below|
MA3-044 detects muscarinic 2 acetylcholine receptor (m2AChR) from human, mouse, pig and rat tissues. This antibody does not detect mAChR from chicken tissues. MA3-044 is specific for the m2 mAChR subtype.
MA3-044 has been successfully used in Western blot and immunoprecipitation protocols. By Western blot, this antibody detects 60-80 kDa protein(s) (depending on the degree of glycosylation) representing the m2AChR from pig heart.
The MA3-044 antigen is purified porcine cardiac m2 AChR.
Acetylcholine receptors (AChRs) mediate synaptic transmission at the neuromuscular junction. The channel-linked AChR that mediates rapid, excitatory actions of acetylcholine is called nicotinic AChR (nAChR) because it can be activated by nicotine. The non-channel linked AChR that medicates the slow actions of acetylcholine, which can be either inhibitory or excitatory, is called muscarinic AChR (mAChR) because it can be activated by muscarine.
The mAChRs are present in neurons of the central and peripheral nervous systems, cardiac and smooth muscle and various exocrine glands. There are 5 subtypes (m1-m5) of the receptor that have a tissue specific pattern of expression. The m2 receptor is localized primarily in cardiac tissue and is also expressed at low levels in the hippocampus, cortex, striatum, thalamus, basal forebrain, brainstem, lung, vas deferens and uterus.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Smooth muscle adaptation and recovery of contractility after massive small bowel resection in rats.
MA3-044 was used in western blot to study muscarinic M2 and M3 receptor expression and smooth muscle contractility during recovery from major small bowel resection
|Chen J,Wen J,Cai W||Experimental biology and medicine (Maywood, N.J.) (237:578)||2012|
Cholesterol as a determinant of cooperativity in the M2 muscarinic cholinergic receptor.
MA3-044 was used in western blot to investigate the role of M2 muscarinic cholinergic receptors
|Colozo AT,Park PS,Sum CS,Pisterzi LF,Wells JW||Biochemical pharmacology (74:236)||2007|
Cooperativity manifest in the binding properties of purified cardiac muscarinic receptors.
MA3-044 was used in western blot to characterize the binding capability of muscarinic receptors
|Wreggett KA,Wells JW||The Journal of biological chemistry (270:22488)||1995|