Anti-CaVα2δ3 (extracellular)-A

Anti-Ca_Vα2δ3 (extracellular)-ATTO-594

Voltage-dependent calcium channel subunit alpha-2/delta-3
Cat #: ACC-103-AR
Sizes: 50 µl
Source: Rabbit
Type: Polyclonal
Applications: IC, IH, LCI
Reactivity: H, M, R

Application key:

CBE- Cell-based ELISA, FC- Flow cytometry, IC- Immunocytochemistry, IE- Indirect ELISA, IFC- Indirect flow cytometry, IH- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

Species reactivity key:

H- Human, M- Mouse, R- Rat

Alomone Labs is pleased to offer a highly specific antibody directed against an extracelluar epitope of rat Ca_Vα2δ3. Anti-Ca_Vα2δ3 (extracellular) antibody (#ACC-103) can be used in western blot immunocytochemistry applications. It has been designed to recognize Ca_Vα2δ3 from rat, human and mouse samples.

We are pleased to offer a new version of this antibody that is directly labeled with an ATTO-594 fluorescent dye. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. The ATTO-594 fluorescent label belongs to the class of Rhodamine dyes and can be used with fluorescent equipment typically optimized to detect Texas Red and Alexa-594. Anti-Ca_Vα2δ3 (extracellular)-ATTO-594 antibody (#ACC-103-AR) has been tested in immunocytochemisry and immunohistochemistry and is specially suited for experiments requiring simultaneous labeling of different markers.

• Applications
• Specifications
• Scientific Background
• Related Products

Live cell imaging / Immunocytochemistry
Rat PC12 cells (1:25). See immunocytochemistry.
Immunohistochemistry

Expression of Ca_Vα2δ3 in rat hippocampus and cortex
Immunohistochemical staining of rat hippocampal CA3 region (A) and rat neocortex (B) using Anti-Ca_Vα2δ3 (extracellular)-ATTO-594 antibody (#ACC-103-AR). In both, A and B, Ca_Vα2δ3 staining (red) appears in pyramidal neurons (arrows). DAPI is used as the counterstain (blue).
Immunocytochemistry

Expression of Ca_Vα2δ3 in PC12 cells.
Immunocytochemical staining of intact living rat PC12 cells. Extracellular staining of cells using Anti-Ca_Vα2δ3 (extracellular)-ATTO-594 antibody (#ACC-103-AR), (1:25).
Immunogen
Peptide CSWWHSDMTAKAQK, corresponding to amino acid residues 942-955 of rat Ca_vα2δ3 (Accession Q8CFG5). Extracellular.

HomologyMouse, human - identical.
PurityAffinity purified on immobilized antigen.
FormulationLyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN₃.
LabelATTO-594. Maximum absorption 601 nm; maximum fluorescence 627 nm. The fluorescence is excited most efficiently in the 580 - 615 nm range. This label is related to the Rhodamine dyes and can be used with filters used to detect Texas Red and Alexa-594.
Standard quality control of each lotWestern blot analysis (unlabeled antibody, #ACC-103), and immunocytochemistry (labeled antibody).
Peptide confirmationConfirmed by amino acid analysis and mass spectrometry.
Storage before reconstitutionThe antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution50 µl double distilled water (DDW).
Antibody concentration after reconstitution1 mg/ml.
Storage after reconstitutionThe reconstituted solution can be stored at 4°C, protected from the light, for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 × g 5 min).
Control antigen storage before reconstitutionLyophilized powder can be stored intact at room temperature for 2 weeks. For longer periods, it should be stored at -20°C.
Control antigen reconstitution100 µl double distilled water (DDW).
Control antigen storage after reconstitution-20ºC.
Preadsorption Control1 µg peptide per 1 µg antibody.
Scientific background

Voltage-gated Ca²⁺ (Ca_V) channels are ubiquitously expressed and function as Ca²⁺ conducting pores in the plasma membrane¹. On the basis of their voltage activation properties, Ca_V channels can be further divided into two broad groups: the low (T-type) and high (L, N, P, Q and R-type) threshold-activated channels². HVA channels are heteromultimers composed of four independently encoded proteins, the pore-forming α1 subunit, which triggers Ca²⁺ flow across the membrane, and the auxiliary subunits α2δ, γ, and β³. The Ca²⁺ channel α2δ subunit is a heavily glycosylated protein that is encoded by a single gene and post-translationally cleaved to yield α2 and δ subunits linked by a disulfide bond with a single transmembrane segment⁴.  The α2δ subunit regulates many functional aspects of Ca²⁺ channels, such as gating, regulating voltage dependent kinetics, and increasing functional channel density on the plasma membrane⁵.

There are four proteins that comprise Ca_Vα2δ: Ca_Vα2δ1, Ca_Vα2δ2, Ca_Vα2δ3 and Ca_Vα2δ4⁶. The Ca_Vα2δ3 subunit is predominantly expressed in neuronal tissue. The Ca_Vα2δ3 subunit regulates all classes of HVA calcium channels. The Caα2δ3 subunits in the nerve terminal function in synaptic morphogenesis and cytoskeletal organization, and that this role is independent of their function in α1 subunit localization and physiology.

Ca_Vα2δ3 is likely to be the primary presynaptic α2δ isoform mediating morphological development of the neuromuscular junction (NMJ), since null alleles have such a large effect on NMJ development and abolish all action-potential evoked transmission⁷. Recent study shows that methylation-dependent transcriptional silencing of Ca_Vα2δ3 may contribute to the metastatic phenotype of breast cancer⁸.

References

1. Catterall, W.A. (2000) Annu. Rev. Cell. Dev. Biol. 16, 521.
2. Qin, N. et al. (2002) Mol. Pharmacol. 62, 485.
3. De Jongh, K.S. et al. (1990)   J. Biol. Chem. 265, 14738.
4. Sipos, I. et al. (2000) Pflug. Arch. 439, 691.
5. Dolphin, A.C. (2009) Curr. Opin. Neurobiol. 19, 237.
6. Cooper C.L. et al. (1987) J. Biol. Chem. 262, 509.
7. Kurshan P.T. (2009) Nat. Neurosci. 12, 1415.
8. Palmieri, C. et al. (2012) Br. J. Cancer 108, 375.