Anti-Syntrophin antibody [1351] (ab11425)
Key features and details
- Mouse monoclonal [1351] to Syntrophin
- Suitable for: ICC/IF, IP, WB, Flow Cyt
- Reacts with: Mouse, Rat, Human
- Isotype: IgG1
Overview
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Product name
Anti-Syntrophin antibody [1351] -
Description
Mouse monoclonal [1351] to Syntrophin -
Host species
Mouse -
Specificity
ab11425 is known to be reactive with the alpha 1, beta 1 and beta 2 subunits of syntrophin. -
Tested Applications & Species
Application Species Flow Cyt HumanICC/IF HumanWB Rat -
Immunogen
Other Immunogen Type corresponding to Syntrophin. Whole purified syntrophin from Torpedo californica electric organ postsynaptic membrane.
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Epitope
ab11425 is directed against an epitope within the PDZ domain of syntrophin. -
Positive control
- In Western Blot, this antibody gave a positive signal in human, mouse and rat skeletal muscle tissue lysates.
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General notes
ab11425 is seen as the "gold standard" for syntrophin assessment.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.05% Sodium azide
Constituents: PBS, 1% BSA -
Concentration information loading...
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Purity
Immunogen affinity purified -
Primary antibody notes
ab11425 is seen as the "gold standard" for syntrophin assessment. -
Clonality
Monoclonal -
Clone number
1351 -
Isotype
IgG1 -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab11425 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
Application | Species |
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Flow Cyt |
Human
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ICC/IF |
Human
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WB |
Rat
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All applications |
Fish
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Application | Abreviews | Notes |
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ICC/IF | (1) |
Use a concentration of 10 µg/ml.
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IP |
Use a concentration of 5 µg/ml.
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WB | (2) |
Use a concentration of 0.2 µg/ml. Detects a band of approximately 58 kDa (predicted molecular weight: 54 kDa).
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Flow Cyt |
Use 0.5µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
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Notes |
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ICC/IF
Use a concentration of 10 µg/ml. |
IP
Use a concentration of 5 µg/ml. |
WB
Use a concentration of 0.2 µg/ml. Detects a band of approximately 58 kDa (predicted molecular weight: 54 kDa). |
Flow Cyt
Use 0.5µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
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Target
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Function
Adapter protein that binds to and probably organizes the subcellular localization of a variety of membrane proteins. May link various receptors to the actin cytoskeleton and the extracellular matrix via the dystrophin glycoprotein complex. Plays an important role in synapse formation and in the organization of UTRN and acetylcholine receptors at the neuromuscular synapse. Binds to phosphatidylinositol 4,5-bisphosphate. -
Tissue specificity
High expression in skeletal muscle and heart. Low expression in brain, pancreas, liver, kidney and lung. Not detected in placenta. -
Involvement in disease
Long QT syndrome 12 (LQT12) [MIM:612955]: A heart disorder characterized by a prolonged QT interval on the ECG and polymorphic ventricular arrhythmias. They cause syncope and sudden death in response to exercise or emotional stress, and can present with a sentinel event of sudden cardiac death in infancy. Note=The disease is caused by mutations affecting the gene represented in this entry. -
Sequence similarities
Belongs to the syntrophin family.
Contains 1 PDZ (DHR) domain.
Contains 2 PH domains.
Contains 1 SU (syntrophin unique) domain. -
Domain
The PH 1 domain mediates the oligomerization in a calcium dependent manner, and the association with the phosphatidylinositol 4,5-bisphosphate.
The PDZ domain binds to the last three or four amino acids of ion channels and receptor proteins. The association with dystrophin or related proteins probably leaves the PDZ domain available to recruit proteins to the membrane.
The SU domain binds calmodulin in a calcium-dependent manner. -
Post-translational
modificationsPhosphorylated by CaM-kinase II. Phosphorylation may inhibit the interaction with DMD. -
Cellular localization
Cell membrane > sarcolemma. Cell junction. Cytoplasm > cytoskeleton. In skeletal muscle, it localizes at the cytoplasmic side of the sarcolemmal membrane and at neuromuscular junctions. - Information by UniProt
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Database links
- Entrez Gene: 6640 Human
- Entrez Gene: 6641 Human
- Entrez Gene: 6645 Human
- SwissProt: Q13424 Human
- SwissProt: Q13425 Human
- SwissProt: Q13884 Human
- SwissProt: Q61234 Mouse
- Unigene: 31121 Human
see all -
Alternative names
- 59 kDa dystrophin-associated protein A1 acidic component 1 antibody
- Alpha-1-syntrophin antibody
- Pro-TGF-alpha cytoplasmic domain-interacting protein 1 antibody
see all
Images
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All lanes : Anti-Syntrophin antibody [1351] (ab11425) at 5 µg/ml
Lane 1 : Human skeletal muscle tissue lysate - total protein (ab29330)
Lane 2 : Skeletal Muscle (Rat) Tissue Lysate
Lane 3 : Skeletal Muscle (Mouse) Tissue Lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 54 kDa
Observed band size: 58 kDa why is the actual band size different from the predicted?
Additional bands at: 26 kDa, 42 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 12 minutes -
Immunocytochemistry/Immunofluorescence analysis of HeLa cells labeling Syntrophin (green) with ab11425 at 1/20. F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue). Cells were fixed with formaldehyde and incubated with the primary antibody overnight at 4°C. A DyLight 488-conjugated secondary antibody was used. 60X magnification. Right - negative control.
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Immunocytochemistry/Immunofluorescence analysis of A2058 cells labeling Syntrophin (green) with ab11425 at 1/20. F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue). Cells were fixed with formaldehyde and incubated with the primary antibody overnight at 4°C. A DyLight 488-conjugated secondary antibody was used. 60X magnification. Right - negative control.
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Immunocytochemistry/Immunofluorescence analysis of 293 cells labeling Syntrophin (green) with ab11425 at 1/20. F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue). Cells were fixed with formaldehyde and incubated with the primary antibody overnight at 4°C. A DyLight 488-conjugated secondary antibody was used. 60X magnification. Right - negative control.
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Overlay histogram showing SH-SY5Y cells stained with ab11425 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab11425, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
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Syntrophin was immunoprecipitated using 0.5mg Mouse Skeletal Muscle tissue lysate, 5µg of Mouse monoclonal to Syntrophin and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Mouse Skeletal Muscle tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab11425.
Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
Band: 58kDa; Syntrophin
Protocols
Datasheets and documents
References (23)
ab11425 has been referenced in 23 publications.
- Patel A et al. Dystrophin R16/17-syntrophin PDZ fusion protein restores sarcolemmal nNOSµ. Skelet Muscle 8:36 (2018). PubMed: 30466494
- Wasala NB et al. Uniform low-level dystrophin expression in the heart partially preserved cardiac function in an aged mouse model of Duchenne cardiomyopathy. J Mol Cell Cardiol 102:45-52 (2017). PubMed: 27908661
- Hakim CH et al. A Five-Repeat Micro-Dystrophin Gene Ameliorated Dystrophic Phenotype in the Severe DBA/2J-mdx Model of Duchenne Muscular Dystrophy. Mol Ther Methods Clin Dev 6:216-230 (2017). PubMed: 28932757
- Wasala NB et al. Genomic removal of a therapeutic mini-dystrophin gene from adult mice elicits a Duchenne muscular dystrophy-like phenotype. Hum Mol Genet 25:2633-2644 (2016). PubMed: 27106099
- Kountz TS et al. Endogenous N-terminal Domain Cleavage Modulates a1D-Adrenergic Receptor Pharmacodynamics. J Biol Chem 291:18210-21 (2016). PubMed: 27382054