Product nameAnti-CACNA1S antibody [1A]
See all CACNA1S primary antibodies
DescriptionMouse monoclonal [1A] to CACNA1S
Tested applicationsSuitable for: ICC/IF, IHC-Fr, IP, WB, IHC-P, Inhibition Assay, ELISA, Flow Cytmore details
Species reactivityReacts with: Mouse, Rat, Rabbit, Guinea pig, Human
Full length native protein (purified) corresponding to Rabbit CACNA1S. Purified from rabbit muscle T-tubule dihydropyridine receptor.
- rat skeletal muscle lysate
Storage instructionsShipped 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 bufferPreservative: 0.05% Sodium azide
Concentration information loading...
PurityProtein A purified
Primary antibody notesVoltage-sensitive calcium channels mediate the entry of calcium into many types of excitable cells and thus play a key role in neurotransmitter release and excitation-contraction (E-C) coupling. The 1,4-dihydropyridines (DHPs) are synthetic organic compounds which can be used to identify the L-type calcium channels that are found in all types of vertebrate muscle, neuronal and neuroendocrine cells. The DHP receptor is part of the L-type calcium channel complex and is thought to be the voltage sensor in E-C coupling. The purified DHP receptor isolated from triads is composed of at least four subunits. The alpha-1 subunit contains the binding site for the DHPs and shows high sequence homology to the voltage gated sodium channel. The alpha-2 subunit is a large glycoprotein associated with the DHP receptor which was first described in skeletal muscle and is also found in high concentrations in other excitable tissues such as cardiac muscle and brain and in low concentrations in most other tissues studied. The other two subunits that co-purify with the DHP receptor are termed beta and gamma.
Our Abpromise guarantee covers the use of ab2862 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 at an assay dependent concentration. PubMed: 21693436|
|IHC-Fr||1/200. PubMed: 21474431|
|IP||Use at an assay dependent concentration.|
|WB||1/500. Detects a band of approximately 200 kDa.|
|Inhibition Assay||Use at an assay dependent concentration.|
|ELISA||Use at an assay dependent concentration.|
|Flow Cyt||Use at an assay dependent concentration.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
FunctionVoltage-sensitive calcium channels (VSCC) mediate the entry of calcium ions into excitable cells and are also involved in a variety of calcium-dependent processes, including muscle contraction, hormone or neurotransmitter release, gene expression, cell motility, cell division and cell death. The isoform alpha-1S gives rise to L-type calcium currents. Long-lasting (L-type) calcium channels belong to the 'high-voltage activated' (HVA) group. They are blocked by dihydropyridines (DHP), phenylalkylamines, benzothiazepines, and by omega-agatoxin-IIIA (omega-Aga-IIIA). They are however insensitive to omega-conotoxin-GVIA (omega-CTx-GVIA) and omega-agatoxin-IVA (omega-Aga-IVA). Calcium channels containing the alpha-1S subunit play an important role in excitation-contraction coupling in skeletal muscle.
Tissue specificitySkeletal muscle specific.
Involvement in diseaseDefects in CACNA1S are the cause of periodic paralysis hypokalemic type 1 (HOKPP1) [MIM:170400]; also designated HYPOPP. HOKPP1 is an autosomal dominant disorder manifested by episodic flaccid generalized muscle weakness associated with falls of serum potassium levels.
Defects in CACNA1S are the cause of malignant hyperthermia susceptibility type 5 (MHS5) [MIM:601887]; an autosomal dominant disorder that is potentially lethal in susceptible individuals on exposure to commonly used inhalational anesthetics and depolarizing muscle relaxants.
Defects in CACNA1S are the cause of susceptibility to thyrotoxic periodic paralysis type 1 (TTPP1) [MIM:188580]. A sporadic muscular disorder characterized by episodic weakness and hypokalemia during a thyrotoxic state. It is clinically similar to hereditary hypokalemic periodic paralysis, except for the fact that hyperthyroidism is an absolute requirement for disease manifestation. The disease presents with recurrent episodes of acute muscular weakness of the four extremities that vary in severity from paresis to complete paralysis. Attacks are triggered by ingestion of a high carbohydrate load or strenuous physical activity followed by a period of rest. Thyrotoxic periodic paralysis can occur in association with any cause of hyperthyroidism, but is most commonly associated with Graves disease.
Sequence similaritiesBelongs to the calcium channel alpha-1 subunit (TC 1.A.1.11) family. CACNA1S subfamily.
DomainEach of the four internal repeats contains five hydrophobic transmembrane segments (S1, S2, S3, S5, S6) and one positively charged transmembrane segment (S4). S4 segments probably represent the voltage-sensor and are characterized by a series of positively charged amino acids at every third position.
The loop between repeats II and III interacts with the ryanodine receptor, and is therefore important for calcium release from the endoplasmic reticulum necessary for muscle contraction.
modificationsPhosphorylation by PKA activates the calcium channel.
- Information by UniProt
- alpha-1 polypeptide antibody
- CAC1S_HUMAN antibody
- CACH1 antibody
Anti-CACNA1S antibody [1A] (ab2862) at 1/2000 dilution + Mouse skeletal muscle microsomes lysate at 10 µg
HRP-conjugated goat anti-mouse IgG, F(ab')2 polyclonal at 1/30000 dilution
Performed under reducing conditions.
Observed band size: 180 kDa why is the actual band size different from the predicted?
Additional bands at: 100 kDa (possible non-specific binding), 36 kDa (possible non-specific binding)
Exposure time: 2 minutes
Blocked with 5% milk for 30 minutes at room temperature. Incubated with the primary antibody diluted in 5% skim milk in TBST for 8 hours at 4°C.
Lanes 1-4 are ab2862 used to detect CACNA1S in lysate prepared from rabbit hearts. Each lane represents a separate heart.
Frozen tissue chunks were pulverized under liquid nitrogen, and homogenized in 10 vol of buffer (0.25 M sucrose, 1 mM EDTA [pH 7.4]). This and all the following procedures took place in the presence of the protease inhibitor cocktail at 4°C or on ice. The homogenate was centrifuged at 3,500 rpm for 10 min to pellet nuclei and debris. The supernatant was centrifuged at 30,000 rpm for 1 hr to pellet the membranes. The membrane pellet was washed with a solution (Tris-HCl 20 mM [pH 7.4], EDTA 1 mM) and referred to as post-nuclear membrane fraction. The post-nuclear membrane fraction was rehomogenized in a Triton-containing lysis buffer (Tris-HCl 20 mM [pH 7.5], NaCl 0.2 M, EDTA 1 mM, Triton X-100 1%) using Dounce grinder. The mixture was centrifuged at 17,000 rpm for 1 hr, and the supernatant (Triton extract) was used for immunoblotting.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) was performed on normal biopsies of deparaffinized human skeletal muscle tissue. Heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then incubated with ab2862 (1:20) 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.
This product has been referenced in:
- Xiyuan Z et al. NO-sGC Pathway Modulates Ca2+Release and Muscle Contraction in Zebrafish Skeletal Muscle. Front Physiol 8:607 (2017). Read more (PubMed: 28878687) »
- Hu H et al. The molecular architecture of dihydropyrindine receptor/L-type Ca2+ channel complex. Sci Rep 5:8370 (2015). WB . Read more (PubMed: 25667046) »