Immunofluorescent analysis of Nicotinic Acetylcholine Receptor using Anti-Nicotinic Acetylcholine Receptor Monoclonal Antibody (88B) (Product# MA3-043) shows staining in Neuro-2a Cells. Nicotinic Acetylcholine Receptor staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Nicotinic Acetylcholine Receptor (Product# MA3-043) at a dilution of 1:100 over night at 4 °C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product# 35503, Goat Anti-Mouse). Images were taken at 60X magnification.
|Tested species reactivity||Amphibian, Chicken, Fish, Human, Mouse, Rat|
|Published species reactivity||Rabbit, Rat, Mouse, Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Purified Torpedo californica acetylcholine receptor.|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunohistochemistry (Frozen) (IHC (F))||1:1,000|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1:5,000|
* 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.
MA3-043 detects nicotinic acetylcholine receptor (nAChR) gamma and delta subunits in torpedo and the delta subunit in mouse, human, rat, chicken and frog. This antibody does not detect the alpha 1 and beta 1 subunits.
MA3-043 has been successfully used in Western blot, immunohistochemistry and immunoprecipitation procedures. By Western blot, this antibody detects a 60 kDa protein representing the gamma subunit and a 65 kDa protein representing the delta subunit from torpedo skeletal muscle homogenates. Under non-reducing conditions the delta subunit migrates mostly as a dimer of ~130 kDa. Immunohistochemical staining of nAChR in rat skeletal muscle with MA3-043 results in strong staining of the motor endplate.
In Immunohistochemical staining, cryostat or permeabilization of the sections is recommended due to the reactivity of this antibody with the cytoplasmic side of the receptor.
The MA3-043 antigen is purified Torpedo californica acetylcholine receptor.
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 nAChRs fall into two subclasses, the muscle-type and the neuronal-type. The muscle-type receptor is a pentameric glycoprotein of five membrane spanning subunits, (2 alpha 1 and quote;s, beta 1, gamma or epsilon and delta) which form a ligand gated ion channel. The neuronal nAChR is also a pentamer, but unlike the muscle-type, is made up of only two general types of subunits, alpha and beta of which multiple subtypes have been described (alpha 2 to alpha 8 and beta 2 to beta 4).
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Expression of the skeletal muscle dystrophin-dystroglycan complex and syntrophin-nitric oxide synthase complex is severely affected in the type 2 diabetic Goto-Kakizaki rat.
MA3-043 was used in western blot to study the reduced expression of the dystrophin-dystroglycan complex and the syntrophin-NOS complex in skeletal muscle of type 2 diabetic rats and the significance for insulin resistance
|Mulvey C,Harno E,Keenan A,Ohlendieck K||European journal of cell biology (84:867)||2005|
Use of continuous-elution gel electrophoresis as a preparative tool for blot overlay analysis.
MA3-043 was used in western blot to evaluate the use of continuous elution gel electrophoresis to isolate proteins for protein-protein interaction studies in blot overlay analysis
|Mulvey C,Ohlendieck K||Analytical biochemistry (319:122)||2003|
Increased expression of the nicotinic acetylcholine receptor in stimulated muscle.
MA3-043 was used in western blot to study the effect of chronic low frequency stimulation on nicotinic acetylcholine receptor expression in skeletal muscle
|O'Reilly C,Pette D,Ohlendieck K||Biochemical and biophysical research communications (300:585)||2003|
Epsilon subunit-containing acetylcholine receptors in myotubes belong to the slowly degrading population.
MA3-043 was used in western blot to study the role of the epsilon subunit in conferring stability on the muscle acetylcholine receptor
|Sala C,O'Malley J,Xu R,Fumagalli G,Salpeter MM||The Journal of neuroscience : the official journal of the Society for Neuroscience (17:8937)||1997|
Association of acetylcholine receptors with peripheral membrane proteins: evidence from antibody-induced coaggregation.
MA3-043 was used in immunocytochemistry to investigate the interaction of acetylcholine receptors with peripheral membrane proteins
|Bloch RJ,Sealock R,Pumplin DW,Luther PW,Froehner SC||The Journal of membrane biology (138:13)||1994|
Dystrophin in a membrane skeletal network: localization and comparison to other proteins.
MA3-043 was used in ELISA to investigate dystrophin's localization, relative abundance and stability in AChR clusters isolated from primary cultures of neonatal rat myotubes
|Dmytrenko GM,Pumplin DW,Bloch RJ||The Journal of neuroscience : the official journal of the Society for Neuroscience (13:547)||1993|
Monoclonal antibody identifies a 200-kDa subunit of the dihydropyridine-sensitive calcium channel.
MA3-043 was used in immunoprecipitation to characterize the dihydropyridine-sensitive calcium channel
|Morton ME,Froehner SC||The Journal of biological chemistry (262:11904)||1987|