Key features and details
- Mouse monoclonal [34C] to Ryanodine Receptor
- Suitable for: ICC/IF, IHC-P, IHC-Fr, IP, WB, Inhibition Assay
- Reacts with: Mouse, Rat, Sheep, Rabbit, Cow, Dog, Human, Pig, Xenopus laevis, Non human primates
- Isotype: IgG1
Product nameAnti-Ryanodine Receptor antibody [34C]
See all Ryanodine Receptor primary antibodies
DescriptionMouse monoclonal [34C] to Ryanodine Receptor
SpecificityDetects Ryanodine Receptor (RyR)-1 and RyR-2 isoforms. In chickens, this antibody detects the alpha, beta and cardiac isoforms. This antibody also detects RyR-3 in mouse cells. In frog, this antibody detects the alpha and beta isoforms. In fish, this antibody detects the alpha isoform. By Western blot, this antibody detects a 565 kDa protein representing RyR from rat skeletal muscle extracts. In non-mammalian vertebrates, a doublet is seen at 565 kDa representing the alpha and beta isoforms of the receptor. Immunohistochemical staining of RyR in chicken brain results in intense staining of cerebellar Purkinje neurons.
Tested applicationsSuitable for: ICC/IF, IHC-P, IHC-Fr, IP, WB, Inhibition Assaymore details
Species reactivityReacts with: Mouse, Rat, Sheep, Rabbit, Cow, Dog, Human, Pig, Xenopus laevis, Non human primates
Predicted to work with: Fish, Amphibian
Other Immunogen Type corresponding to Chicken Ryanodine Receptor. Partially purified chicken pectoral muscle ryanodine receptor.
- rat skeletal muscle
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -80°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.05% Sodium azide
Concentration information loading...
PurityProtein A purified
Primary antibody notesThe Ryanodine Receptor (RyR) is the channel responsible for calcium release from muscle cell Sarcoplasmic Reticulum (SR) and also plays a role in calcium regulation in non-muscle cells. The RyR exists as a homotetramer and is predicted to have a short cytoplasmic C-terminus and 4-10 transmembrane domains. The remainder of the protein, termed the "foot" region, is located in the cytoplasm between the transverse tubule and the SR. Mammalian RyR isoforms are the product of three different genes: RyR-1 is expressed predominantly in skeletal muscle and areas of the brain; RyR-2 is expressed predominantly in heart muscle but also found in the stomach, endothelial cells and diffuse areas of the brain; and RyR-3 is found in smooth muscle and the brain (striatum, thalamus and hippocampus). In non-mammalian vertebrates, the RyR isoforms are termed alpha, beta and cardiac which correlate loosely to the mammalian RyR-1, RyR-3 and RyR-2 isoforms respectively.
Our Abpromise guarantee covers the use of ab2868 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 1 µg/ml.|
|IHC-Fr||1/100 - 1/1000.|
|IP||Use at an assay dependent concentration.|
|WB||1/5000. Predicted molecular weight: 565 kDa. As RyR is a large protein, we recommend using a 6-8% gel, wet transfering protein overnight at low voltage, adding 0.1% SDS to transfer buffer and reducing methanol to 10% or less.|
|Inhibition Assay||Use at an assay dependent concentration.|
FunctionCalcium channel that mediates the release of Ca(2+) from the sarcoplasmic reticulum into the cytoplasm and thereby plays a key role in triggering muscle contraction following depolarization of T-tubules. Repeated very high-level exercise increases the open probability of the channel and leads to Ca(2+) leaking into the cytoplasm. Can also mediate the release of Ca(2+) from intracellular stores in neurons, and may thereby promote prolonged Ca(2+) signaling in the brain. Required for normal embryonic development of muscle fibers and skeletal muscle. Required for normal heart morphogenesis, skin development and ossification during embryogenesis.
Tissue specificitySkeletal muscle and brain (cerebellum and hippocampus).
Involvement in diseaseMalignant hyperthermia 1
Central core disease of muscle
Multiminicore disease with external ophthalmoplegia
Myopathy, congenital, with fiber-type disproportion
Defects in RYR1 may be a cause of Samaritan myopathy, a congenital myopathy with benign course. Patients display severe hypotonia and respiratory distress at birth. Unlike other congenital myopathies, the health status constantly improves and patients are minimally affected at adulthood.
Sequence similaritiesBelongs to the ryanodine receptor (TC 1.A.3.1) family. RYR1 subfamily.
Contains 3 B30.2/SPRY domains.
Contains 5 MIR domains.
DomainThe calcium release channel activity resides in the C-terminal region while the remaining part of the protein constitutes the 'foot' structure spanning the junctional gap between the sarcoplasmic reticulum (SR) and the T-tubule.
modificationsChannel activity is modulated by phosphorylation. Phosphorylation at Ser-2843 may increase channel activity. Repeated very high-level exercise increases phosphorylation at Ser-2843.
Activated by reversible S-nitrosylation. Repeated very high-level exercise increases S-nitrosylation.
Cellular localizationSarcoplasmic reticulum membrane. Membrane. The number of predicted transmembrane domains varies between orthologs, but both N-terminus and C-terminus seem to be cytoplasmic.
- Information by UniProt
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Representative Western blots for Ser(P)2814 and Ryanodine Receptor in control and Cav1.2I/E mutant mouse hearts. Ryanodine Receptor (bottom panel) was detected with ab2868.
ICC/IF image of ab2868 stained PC12 cells. The cells were 4% formaldehyde 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 (ab2868, 1µg/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.
ab2868 staining Ryanodine Receptor (red) in Mouse Skeletal muscle cells at day 10 of agrin-treated differentation by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.25% Triton X-100 in PBS and blocked with 10% serum for 45 minutes at 20°C. Samples were incubated with primary antibody (1/200 in PBS + 3% BSA) for 2 hours at 20°C. An Alexa Fluor® 647-conjugated Donkey anti-mouse IgG polyclonal (1/500) was used as the secondary antibody. Nucleus stained blue.
Immunohistochemistry was performed on biopsies of deparaffinized Human skeletal muscle tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer 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 Ryanodine Receptor ab2868 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.
ab2868 at a 1/200 dilution staining Ryanodine Receptor in mouse liver tissue sections by Immunohistochemistry (frozen sections) incubated for 9 hours at +4°C. Fixed in formaldehyde, permeabilized using 0.2% Triton X-100. Blocked using 2% BSA for 30 minutes at 20°C. Secondary used at a 1/200 dilution polyclonal Goat anti-mouse IgG conjugated to Alexa Fluor® 555.
ab2868 staining Ryanodine Receptor in Rat cardiomyocyte cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with methanol/acetone (1:1) and blocked with 3% BSA for 1 hour at 18°C. Samples were incubated with primary antibody (1/300 in PBS + 3% BSA) for 16 hours at 4°C. An Alexa Fluor® 546-conjugated Goat anti-mouse IgG (H+L) polyclonal (1/300) was used as the secondary antibody.
ab2868 has been referenced in 48 publications.
- Zügel M et al. Moderate intensity continuous training reverses the detrimental effects of ovariectomy on RyR1 phosphorylation in rat skeletal muscle. Mol Cell Endocrinol 481:1-7 (2019). PubMed: 30465874
- Estañ MC et al. Recessive mutations in muscle-specific isoforms of FXR1 cause congenital multi-minicore myopathy. Nat Commun 10:797 (2019). PubMed: 30770808
- Aloni E et al. Synaptopodin Deficiency Ameliorates Symptoms in the 3xTg Mouse Model of Alzheimer's Disease. J Neurosci 39:3983-3992 (2019). PubMed: 30872324
- Lotteau S et al. A Mechanism for Statin-Induced Susceptibility to Myopathy. JACC Basic Transl Sci 4:509-523 (2019). PubMed: 31468006
- Chen J et al. Panax Notoginseng Saponins Protect Cardiac Myocytes Against Endoplasmic Reticulum Stress and Associated Apoptosis Through Mediation of Intracellular Calcium Homeostasis. Front Pharmacol 10:1013 (2019). PubMed: 31616293