Guinea pig Anti-CaV1.2

Guinea pig Anti-Ca_V1.2

α_1c, L-type of Voltage-Gated Ca²⁺ Channel, Cacna1c
Cat #: AGP-001
Sizes: 25 µl, 50 µl, 0.2 ml
Source: Guinea pig
Type: Polyclonal
Applications: IH, WB
May also work in: IC, IFC, IP
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
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Immuno-colocalization of Ca_V1.2 and GABA(A) α1 Receptor in rat and mouse hippocampusImmunohistochemical staining of mouse and rat hippocampal dentate gyrus using Guinea pig Anti-Ca_V1.2 antibody (#AGP-001) and Anti-GABA(A) α1 Receptor (extracellular)-ATTO-488 antibody (#AGA-001-AG) in the same section. Both Ca_V1.2 (red) and GABA(A) α1 Receptor (green) are detected in neuron-shapep cells (arrows). Staining suggests partial colocalization between Ca_V1.2 and GABA(A) α1 Receptor in a sub-population of dentate gyrus neurons. A. Mouse hippocampus. B. Rat hippocampus.

Guinea pig Anti-Ca_V1.2 (#AGP-001), raised in guinea pigs, is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot and immunohistochemistry applications. It and has been designed to recognize Ca_V1.2 from mouse, rat and human samples. The antigen used to immunize guinea pigs is the same as Anti-Ca_V1.2 antibody (#ACC-003) raised in rabbit. Our line of guinea pig antibodies enables more flexibility with our products such as immuno-colocalization studies, immunoprecipitation, etc.

For a list of product citations in the literature, see product citations tab below. If you know of additional relevant citations for this product, please let us know.

• Applications
• Specifications
• Scientific Background
• Citations
• Related Products

Western blot

Western blot analysis of rat brain membrane:
1. Guinea pig Anti-Ca_v1.2 antibody (#AGP-001), (1:200).
2. Anti-Ca_v1.2 antibody, preincubated with the control peptide antigen.

Western blot analysis of Ca_V1.2-transfected Xenopus oocytes (lane 1) and non-transfected oocytes lysates (lane 2):
1. Guinea pig Anti-Ca_V1.2 antibody (#AGP-001), (1:200) in Ca_V1.2 Channel Membrane Lysate (LX-104).
2. Anti-Cav1.2 antibody in non-transfected oocytes.
Immunohistochemistry

Immuno-colocalization of Ca_V1.2 and GABA(A) α1 Receptor in rat cerebellum
Immunohistochemical staining of rat cerebellum using Guinea pig Anti-Ca_V1.2 antibody (#AGP-001) and Anti-GABA(A) α1 Receptor (extracellular)-ATTO-488 antibody (#AGA-001-AG). A. Ca_V1.2 (red) is detected mostly in Purkinje cells (arrow). B. In the same section, GABA(A) α1 Receptor (green) is observed in the granule layer. C. Merge of the two images suggests some colocalization between Ca_V1.2 and GABA(A) α1 Receptor in the rat granule layer but only Ca_V1.2 appears in Purkinje cells.

Expression of Ca_V1.2 in human atria
Immunohistochemical staining of human left atrium using Guinea pig Anti-Ca_V1.2 antibody (#AGP-001), (1:100).

The picture was kindly provided by Dr. Van Wagoner, D.R. from the Department of Molecular Cardiology, Cleveland Clinic, Cleveland, Ohio, USA. Lovano, B. and Peterson, J. collected the data.

Expression of Ca_v1.2 in rat heart
Immunohistochemical staining of rat heart paraffin embedded sections using Guinea pig Anti-Ca_v1.2 antibody (#AGP-001). A. Ca_v1.2 staining (green) appears mainly in the cardiac muscle, and in a lesser intensity in the tunica intima layer of the smooth muscle of the muscular arteries. B. Nuclear staining using DAPI as the counter stain. C. Merged images of A and B.

Expression of Ca_V1.2 in mouse hippocampus
Immunohistochemical staining of mouse dentate gyrus using Guinea pig Anti-Ca_V1.2 antibody (#AGP-001). A. Ca_V1.2 (green) appeared in the outer molecular layer of the dentate gyrus and in the granule layer. B. Counterstain with DAPI (blue) outlines the granule layer of the dentate gyrus.
Immunogen
Peptide (C)TTKINMDDLQPSENEDKS, corresponding to amino acid residues 848-865 of rat Ca_v1.2 (Accession P22002). Intracellular loop between domains II and III.

HomologyMouse - identical; guinea pig -17/18 amino acid residues identical; human, rabbit - 16/18 amino acid residues identical.
PurityAffinity purified on immobilized antigen.
FormulationLyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN₃.
Standard quality control of each lotWestern blot analysis.
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.
Reconstitution25 µl, 50 µl or 0.2 ml double distilled water (DDW), depending on the sample size.
Antibody concentration after reconstitution0.8 mg/ml.
Storage after reconstitutionThe reconstituted solution can be stored at 4°C 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 x 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⁺ channels (Ca_V), enable the passage of Ca²⁺ ions in a voltage dependent manner. These heteromeric entities are formed in part by the pore-forming α1 subunit which determines the biophysical and pharmacological properties of the channel¹.

L-type Ca²⁺ channels make up one of three voltage-gated Ca²⁺ channel families. Four different α1 isoforms (Ca_V1.1 to Ca_V1.4) belong to the L-type subfamily. Structurally, each α1 subunit has four homologous domains (I-IV) and each domain has a six transmembrane section. Like many other voltage-gated channels, L-type Ca²⁺ channels have auxiliary subunits which are responsible for modulating the surface expression and properties of the channels^2-5.

Ca_V1.1 is mostly expressed in the skeletal muscle, while Ca_V1.4 is mainly detected in the retina. The expression of both Ca_V1.2 and Ca_V1.3 is more extensive and includes neurons, heart, smooth muscle, inner ear, retina and pancreas⁶. L-type Ca²⁺ channels are involved in and modulate a variety of physiological functions such as muscle contraction, hormone secretion, neuronal excitability and gene expression⁵.

Ca_V1.2 undergoes various post-translational modifications. For example, it can undergo proteolytic cleavage at its C-terminal. This cleavage has been shown to take place in neurons following the activation of NMDA receptors^5,7 and in the heart^5,8,9. The cleaved moiety can still interact with the channel and its general purpose is to modulate channel activity⁵. Other postranslation modifications of the channel include phosphorylation of Ca_V1.2 by a number of kinases such as PKA, PKC, Src and CaMKII⁵. In addition, it is not surprising that phosphatases also regulate channel activity, as they are required to antagonize the activity of the various kinases known to phosphorylate Ca_V1.2 ⁵.

The fact that Ca_V1.2 plays a prominent role in proper cardiac function has prompted endless studies regarding its regulation. Such studies have concluded that dysregulation of the channel leads to anomalies in heart contraction and thus heart failure⁵. Likewise, Ca_V1.2 defects have been detected in autism and bipolar disorder¹⁰.

References

1. Bauer, C.S. et al. (2010) Curr. Opin. Neurobiol. 20, 563.
2. Arikkath, J. et al. (2003) Curr. Opin. Neurobiol. 13, 298.
3. Catterall, W.A. (2000) Annu. Rev. Cell. Dev. Biol. 16, 521.
4. Davies, A. et al. (2007) Trends Pharmacol. Sci. 28, 220.
5. Dai, S. et al. (2009) Physiol. Rev. 89, 411.
6. Zuccotti, A. et al. (2011) Trends Pharmacol. Sci. 32, 366.
7. Hell, J.W. et al. (1996) Proc. Natl. Acad. Sci. U.S.A. 93, 3362.
8. De Jongh, K.S. et al. (1996) Biochemistry 35, 10392.
9. Gao, T. et al. (2001) J. Biol. Chem. 276, 21089.
10. Liao, P. et al. (2010) Pflugers. Arch. 460, 353.