The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration. Detects a band of approximately 190 kDa (predicted molecular weight: 268 kDa).Can be blocked with Rat CACNA1A peptide (ab32641). PubMed: 21293071
CACNA1a has a major form with an apparent molecular mass of 190 kDa and a minor, full-length form with an apparent molecular mass of 220 kDa. [PMID:7673157]
Cav2.1 is a voltage-sensitive calcium channels (VSCC) which belongs to the calcium channel alpha-1 subunit family. Cav2.1 mediates the entry of calcium ions into excitable cells and is 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. Cav2.1 (isoform alpha-1A) gives rise to P and/or Q-type calcium currents. Voltage-dependent calcium channels are multisubunit complexes, consisting of alpha-1, alpha-2, beta and delta subunits in a 1:1:1:1 ratio. The channel activity is directed by the pore-forming and voltage-sensitive alpha-1 subunit. In many cases, this subunit is sufficient to generate voltage-sensitive calcium channel activity. The auxiliary subunits beta and alpha-2/delta linked by a disulfide bridge regulate the channel activity.
This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab32642 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
Western blot - Anti-CACNA1A antibody (ab32642)
Anti-CACNA1A antibody (ab32642) at 1 µg/ml + Cerebellum Rat Tissue Lysate at 20 µg
Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-CACNA1A antibody (ab32642)This image is courtesy of Sophie Pezet, Laboratoire de Neurobiologie, Paris, France
Immunofluorescent staining for CACNA1A using Rabbit polyclonal to CACNA1A (ab32642)in rat Purkinje (arrows) and granular (arrow heads) cells, of the rat cerebellum.
Protocol details: Rats were intracardially perfused with 4% paraformaldehyde. Whole brain tissue was post-fixed overnight in the same fixative, and cryoprotected in 20% sucrose and frozen in OCT. 30µm coronal sections were cut by cyrostat for use in fre floating IHC. Primary antibody ab32642 was incubated overnight at 1/3000 at room temperature. Secondary antibody Alexa fluor 488 1/3000 was incubated for 2 hours at room temperature.
Grauel MK et al. RIM-binding protein 2 regulates release probability by fine-tuning calcium channel localization at murine hippocampal synapses. Proc Natl Acad Sci U S A113:11615-11620 (2016).
Read more (PubMed: 27671655) »
Vares G et al. Gene silencing of Tead3 abrogates radiation-induced adaptive response in cultured mouse limb bud cells. J Radiat Res (Tokyo)52:39-46 (2011).
Read more (PubMed: 21293071) »